View source for Template:Team:Groningen/CONTENT/EXPERIMENTS/Overproduce BslA in B. subtilis ComI

{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 16 April 2015 - 00:00, 16 April 2015
	|executor=Atze van Stralen
	|content=

{{Template:Team:Groningen/TEMPLATES/OBJECT/FIGURE
	|type=small
	|imgversion=/b/be
	|image=primers_bslA_0001b
	|caption=Primers.
}}<html>
<div class="text">
The following primers were designed for bslA (the gene responsible for the 
production of BslA) in B. subtilis 168. 
</div>
<div class="object data" id="tbl1">
	<div class="wrapper">
	<div class="header">
		<div class="field fw1">#</div>
		<div class="field fw3">Name</div>
		<div class="field fwx">Sequence</div>
	</div>
	<div class="record">
		<div class="field fw1">1</div>
		<div class="field fw3">bslA for</div>
		<div class="field 
fwx">GCGAATTCGCGGCCGCTTCTAGAGCATTTTTTAGGGGGAATTTTGTTATG</div>
	</div>
	<div class="record">
		<div class="field fw1">2</div>
		<div class="field fw3">bslA overlap for</div>
		<div class="field 
fwx">GATTTGTTAGGAGCTGGCGAATTTAAATTAAAACTG</div>
	</div>
	<div class="record">
		<div class="field fw1">3</div>
		<div class="field fw3">bslA overlap rev</div>
		<div class="field 
fwx">CAGTTTTAATTTAAATTCGCCAGCTCCTAACAAATC</div>
	</div>
	<div class="record">
		<div class="field fw1">4</div>
		<div class="field fw3">bslA rev</div>
		<div class="field 
fwx">CGTACTAGTAGCGGCCGCTGCAGTTATTAGTTGCAACCGCAAGGCTGAG</div>
	</div>
	</div>
	<div class="caption">Primer design for bslA. </div>
</div>
</html>
}}

{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 13 May 2015 - 00:00, 13 May 2015
	|executor=Harm Ruesink
	|content=<html>
<div class="subtitle">PCR 1</div>
<div class="text">Prepare dNTP Mastermix with final concentration of 2 mM of 
each dNTP. </div>
<div class="object data" id="tbl1">
	<div class="wrapper">
	<div class="header">
		<div class="field fw3">Ingredient</div>
		<div class="field fw3">Amount</div>
	</div>
	<div class="record">
		<div class="field fw3">100 mM dATP</div>
		<div class="field fw3">20 µL</div>
	</div>
	<div class="record">
		<div class="field fw3">100 mM dTTP</div>
		<div class="field fw3">20 µL</div>
	</div>
	<div class="record">
		<div class="field fw3">100 mM dCTP</div>
		<div class="field fw3">20 µL</div>
	</div>
	<div class="record">
		<div class="field fw3">100 mM dGTP</div>
		<div class="field fw3">20 µL</div>
	</div>
	<div class="record">
		<div class="field fw3">\( \mathrm{H_2 O} \)</div>
		<div class="field fw3">920 µL</div>
	</div>
	</div>
	<div class="caption">dNTP Mastermix.</div>
</div>
</html>
}}
{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 13 May 2015 - 00:00, 13 May 2015
	|executor=Harm Ruesink
	|content=<html>
<div class="text">Per sample:</div>
<div class="object data" id="tbl1">
	<div class="wrapper">
	<div class="header">
		<div class="field fw5">Ingredient</div>
		<div class="field fw2">Amount</div>
	</div>
	<div class="record">
		<div class="field fw5">Mastermix PCR 1</div>
		<div class="field fw2">30 µL</div>
	</div>
	<div class="record">
		<div class="field fw5">template DNA (B. subtilis 168 DNA)</div>
		<div class="field fw2">0.3 µL</div>
	</div>
	<div class="record">
		<div class="field fw5">bslA for</div>
		<div class="field fw2">0.3 µL</div>
	</div>
	<div class="record">
		<div class="field fw5">bslA overlap for</div>
		<div class="field fw2">0.3 µL</div>
	</div>
	<div class="record">
		<div class="field fw5">bslA overlap rev</div>
		<div class="field fw2">0.3 µL</div>
	</div>
	<div class="record">
		<div class="field fw5">bslA rev</div>
		<div class="field fw2">0.3 µL</div>
	</div>
	</div>
	<div class="caption">Mastermix PCR 1.</div>
</div>
<div class="subtitle"> Samples</div>
<div class="object data" id="tbl1">
	<div class="wrapper">
	<div class="header">
		<div class="field fw1">#</div>
		<div class="field fwx">Description</div>
	</div>
	<div class="record">
		<div class="field fw1">1</div>
		<div class="field fwx">yuaB for + yuaB overlap rev</div>
	</div>
	<div class="record">
		<div class="field fw1">2</div>
		<div class="field fwx">yuaB overlap for + yuaB rev</div>
	</div>
	</div>
	<div class="caption">Samples.</div>
</div>
<div class="subtitle"> PCR thermocycle </div>
<div class="object data" id="tbl1">
	<div class="wrapper">
	<div class="header">
		<div class="field fw1">#</div>
                <div class="field fw5">Step</div>
		<div class="field fw2">Temperature</div>
                <div class="field fw2">Time</div>
	</div>
	<div class="record">
		<div class="field fw1">1</div>
		<div class="field fw5">Initial denaturation</div>
                <div class="field fw2">98 °C</div>
                <div class="field fw2">2:00</div>
	</div>
	<div class="record">
		<div class="field fw1">2</div>
		<div class="field fw5">Denaturation</div>
                <div class="field fw2">98 °C</div>
                <div class="field fw2">0:10</div>
	</div>
	<div class="record">
		<div class="field fw1">3</div>
		<div class="field fw5">Annealing</div>
                <div class="field fw2">60 °C</div>
                <div class="field fw2">0:20</div>
	</div>
	<div class="record">
		<div class="field fw1">4</div>
		<div class="field fw5">Extension</div>
                <div class="field fw2">72 °C</div>
                <div class="field fw2">0:30</div>
	</div>
	<div class="record">
		<div class="field fw1">5</div>
		<div class="field fw5">Go back to 2 (repeat 30x)</div>
                <div class="field fw2"></div>
                <div class="field fw2"></div>
	</div>
	<div class="record">
		<div class="field fw1">6</div>
		<div class="field fw5">Final extension</div>
                <div class="field fw2">72 °C</div>
                <div class="field fw2">5:00</div>
	</div>
	</div>
	<div class="caption">Recommended thermocycle.</div>
</div> 
<div class="subtitle">Gel results</div>
<div class="text">
Sample 1 formed no product. Sample 2 formed a product of ±200bp visible on gel. 
Sample 2 was purified using PCR and Stored in -20°C as 
"1.2". 
</div>
</html>
}}

{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 15 May 2015 - 00:00, 15 May 2015
	|executor=Harm Ruesink
	|content=<html>
<div class="subtitle">PCR 2</div>
<div class="text"> The following samples were prepared using B. subtilis 168 
DNA.</div>
<div class="object data" id="tbl1">
	<div class="wrapper">
	<div class="header">
		<div class="field fw3">Ingredient</div>
		<div class="field fw3">Quantity</div>
	</div>
	<div class="record">
		<div class="field fw3">5x buffer</div>
		<div class="field fw3">40 µL</div>
	</div>
	<div class="record">
		<div class="field fw3">dNTP MM (2mM)</div>
		<div class="field fw3">2 µL</div>
	</div>
	<div class="record">
		<div class="field fw3">Phusion polymerase</div>
		<div class="field fw3">20 µL</div>
	</div>
	<div class="record">
		<div class="field fw3">\( \mathrm{\mathrm{H_2 O}} \)</div>
		<div class="field fw3">136 µL</div>
	</div>
	</div>
	<div class="caption">Mastermix PCR 2.</div>
</div>
<div class="text">Per sample:</div>
<div class="object data" id="tbl1">
	<div class="wrapper">
	<div class="header">
		<div class="field fw3">Ingredient</div>
		<div class="field fw3">Quantity</div>
	</div>
	<div class="record">
		<div class="field fw3">Mastermix PCR 2</div>
		<div class="field fw3">30 µL</div>
	</div>
	<div class="record">
		<div class="field fw3">Per template DNA</div>
		<div class="field fw3">2 µL</div>
	</div>
	</div>
	<div class="caption">Mix per sample.</div>
</div>
<div class="text"> A sample marked "4" was prepared with the following primers: 
yuaB for + yuaB overlap rev. Template DNA was B. subtilis 168 DNA. The 
following 
thermocycle was performed.</div>

<div class="object data" id="tbl1">
	<div class="wrapper">
	<div class="header">
		<div class="field fw1">#</div>
		<div class="field fw5">Step</div>
		<div class="field fw2">Temperature</div>
                <div class="field fw2">Time</div>
	</div>
	<div class="record">
		<div class="field fw1">1</div>
		<div class="field fw5">Initial denaturation</div>
		<div class="field fw2">98 °C</div>
                <div class="field fw2">3:00</div>
	</div>
	<div class="record">
		<div class="field fw1">2</div>
		<div class="field fw5">Denaturation</div>
		<div class="field fw2">98 °C</div>
                <div class="field fw2">0:30</div>
	</div>
	<div class="record">
		<div class="field fw1">3</div>
		<div class="field fw5">Annealing</div>
		<div class="field fw2">62 °C</div>
                <div class="field fw2">0:30</div>
	</div>
	<div class="record">
		<div class="field fw1">4</div>
		<div class="field fw5">Extention</div>
		<div class="field fw2">72°C</div>
                <div class="field fw2">1:00</div>
	</div>
	<div class="record">
		<div class="field fw1">5</div>
		<div class="field fw5">Go back to step 2 (repeat 3x)</div>
		<div class="field fw2"></div>
                <div class="field fw2"></div>
	</div>
	<div class="record">
		<div class="field fw1">6</div>
		<div class="field fw5">Denaturation</div>
		<div class="field fw2">98 °C</div>
                <div class="field fw2">0:30</div>
	</div>
	<div class="record">
		<div class="field fw1">7</div>
		<div class="field fw5">Annealing</div>
		<div class="field fw2">60 °C</div>
                <div class="field fw2">0:30</div>
	</div>
	<div class="record">
		<div class="field fw1">8</div>
		<div class="field fw5">Extention</div>
		<div class="field fw2">72 °C</div>
                <div class="field fw2">1:00</div>
	</div>
	<div class="record">
		<div class="field fw1">9</div>
		<div class="field fw5">Go back to step 6 (repeat 3x) </div>
		<div class="field fw2"></div>
                <div class="field fw2"></div>
	</div>
	<div class="record">
		<div class="field fw1">10</div>
		<div class="field fw5">Denaturation</div>
		<div class="field fw2">98 °C</div>
                <div class="field fw2">0:30</div>
	</div>
	<div class="record">
		<div class="field fw1">11</div>
		<div class="field fw5">Annaeling</div>
		<div class="field fw2">58°C</div>
                <div class="field fw2">0:30</div>
	</div>
	<div class="record">
		<div class="field fw1">12</div>
		<div class="field fw5">Extention</div>
		<div class="field fw2">72 °C</div>
                <div class="field fw2">1:00</div>
	</div>
	<div class="record">
		<div class="field fw1">13</div>
		<div class="field fw5">Go back to step 10 (repeat 3x)</div>
		<div class="field fw2"></div>
                <div class="field fw2"></div>
	</div>
	<div class="record">
		<div class="field fw1">14</div>
		<div class="field fw5">Denaturation</div>
		<div class="field fw2">98 °C</div>
                <div class="field fw2">0:30</div>
	</div>
	<div class="record">
		<div class="field fw1">15</div>
		<div class="field fw5">Annaaling</div>
		<div class="field fw2">56 °C</div>
                <div class="field fw2">0:30</div>
	</div>
	<div class="record">
		<div class="field fw1">16</div>
		<div class="field fw5">Extention</div>
		<div class="field fw2">72 °C</div>
                <div class="field fw2">1:00</div>
	</div>
	<div class="record">
		<div class="field fw1">17</div>
		<div class="field fw5">Go back to step 14 (repeat 3x)</div>
		<div class="field fw2"></div>
                <div class="field fw2"></div>
	</div>
	<div class="record">
		<div class="field fw1">18</div>
		<div class="field fw5">Denaturation</div>
		<div class="field fw2">98 °C</div>
                <div class="field fw2">0:30</div>
	</div>
	<div class="record">
		<div class="field fw1">19</div>
		<div class="field fw5">Annealing</div>
		<div class="field fw2">54 °C</div>
                <div class="field fw2">0:30</div>
	</div>
	<div class="record">
		<div class="field fw1">20</div>
		<div class="field fw5">Extention</div>
		<div class="field fw2">72 °C</div>
                <div class="field fw2">1:00</div>
	</div>
	<div class="record">
		<div class="field fw1">21</div>
		<div class="field fw5">Go back to step 18 (repeat 3x)</div>
		<div class="field fw2"></div>
                <div class="field fw2"></div>
	</div>
	<div class="record">
		<div class="field fw1">22</div>
		<div class="field fw5">Denaturation</div>
		<div class="field fw2">98 °C</div>
                <div class="field fw2">0:30</div>
	</div>
	<div class="record">
		<div class="field fw1">23</div>
		<div class="field fw5">Annealing</div>
		<div class="field fw2">54 °C</div>
                <div class="field fw2">0:30</div>
	</div>
	<div class="record">
		<div class="field fw1">24</div>
		<div class="field fw5">Extention</div>
		<div class="field fw2">72 °C</div>
                <div class="field fw2">1:00</div>
	</div>
	<div class="record">
		<div class="field fw1">25</div>
		<div class="field fw5">Go back to step 22 (repeat 30x)</div>
		<div class="field fw2"></div>
                <div class="field fw2"></div>
	</div>
	<div class="record">
		<div class="field fw1">26</div>
		<div class="field fw5">Final Extention</div>
		<div class="field fw2">72 °C</div>
                <div class="field fw2">10:00</div>
	</div>
	</div>
	<div class="caption">Recommended thermocycle.</div>
</div>


</html>
}}


{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 18 May 2015 - 00:00, 18 May 2015
	|executor=Harm Ruesink
	|content=<html>
<div class="subtitle">PCR 3</div>
<div class="object data" id="tbl1">
	<div class="wrapper">
	<div class="header">
		<div class="field fw3">Ingredient</div>
		<div class="field fw3">Quantity</div>
	</div>
	<div class="record">
		<div class="field fw3">5x buffer</div>
		<div class="field fw3">28 µL</div>
	</div>
	<div class="record">
		<div class="field fw3">dNTP MM (2mM)</div>
		<div class="field fw3">14 µL</div>
	</div>
	<div class="record">
		<div class="field fw3">Phusion polymerase</div>
		<div class="field fw3">1.4 µL</div>
	</div>
	<div class="record">
		<div class="field fw3">\( \mathrm{H_2 O} \)</div>
		<div class="field fw3">95.2 µL</div>
	</div>
	</div>
	<div class="caption">Mastermix PCR 3.</div>
</div>
<div class="text">Per sample:</div>
<div class="object data" id="tbl1">
	<div class="wrapper">
	<div class="header">
		<div class="field fw3">Ingredient</div>
		<div class="field fw3">Quantity</div>
	</div>
	<div class="record">
		<div class="field fw3">Mastermix PCR 3</div>
		<div class="field fw3">30 µL</div>
	</div>
	<div class="record">
		<div class="field fw3">Per template DNA</div>
		<div class="field fw3">0.3 µL</div>
	<div class="record">
		<div class="field fw3">Per primer</div>
		<div class="field fw3">0.3 µL</div>
	</div>
	</div>
	</div>
	<div class="caption">Mix per sample.</div>
</div>
<div class="text">The following samples were prepared: 1. primers: yuaB for + 
yuaB rev, template DNA: B. subtilis 168 DNA 
2. primers: yuaB for + yuaB overlap rev, template DNA: B. subtilis 168 DNA 
</div>
<div class="object data" id="tbl1">
	<div class="wrapper">
	<div class="header">
		<div class="field fw1">#</div>
		<div class="field fw5">Primers</div>
		<div class="field fw5">Template DNA</div>
	</div>
	<div class="record">
		<div class="field fw1">1</div>
		<div class="field fw5">yuaB for + yuaB rev</div>
		<div class="field fw5">B. subtilis 168 DNA</div>
	</div>
	<div class="record">
		<div class="field fw1">2</div>
		<div class="field fw5">yuaB for + yuaB overlap rev</div>
		<div class="field fw5">B. subtilis 168 DNA</div>
	</div>
	</div>
	<div class="caption"> </div>
</div>

<div class="object data" id="tbl1">
	<div class="wrapper">
	<div class="header">
		<div class="field fw1">#</div>
		<div class="field fw5">Step</div>
		<div class="field fw2">Temperature</div>
                <div class="field fw2">Time</div>
	</div>
	<div class="record">
		<div class="field fw1">1</div>
		<div class="field fw5">Initial denaturation</div>
		<div class="field fw2">98 °C</div>
                <div class="field fw2">3:00</div>
	</div>
	<div class="record">
		<div class="field fw1">2</div>
		<div class="field fw5">Denaturation</div>
		<div class="field fw2">98 °C</div>
                <div class="field fw2">0:30</div>
	</div>
	<div class="record">
		<div class="field fw1">3</div>
		<div class="field fw5">Annealing</div>
		<div class="field fw2">56 °C</div>
                <div class="field fw2">0:30</div>
	</div>
	<div class="record">
		<div class="field fw1">4</div>
		<div class="field fw5">Extention</div>
		<div class="field fw2">72 °C</div>
                <div class="field fw2">1:00</div>
	</div>
	<div class="record">
		<div class="field fw1">5</div>
		<div class="field fw5">Go back to step 6(repeat 15x)</div>
		<div class="field fw2"></div>
                <div class="field fw2"></div>
	</div>
	<div class="record">
		<div class="field fw1">6</div>
		<div class="field fw5">Denaturation</div>
		<div class="field fw2">98 °C</div>
                <div class="field fw2">0:30</div>
	</div>
	<div class="record">
		<div class="field fw1">7</div>
		<div class="field fw5">Annealing</div>
		<div class="field fw2">70 °C</div>
                <div class="field fw2">0:30</div>
	</div>
	<div class="record">
		<div class="field fw1">8</div>
		<div class="field fw5">Extention</div>
		<div class="field fw2">72 °C</div>
                <div class="field fw2">1:00</div>
	</div>
	<div class="record">
		<div class="field fw1">9</div>
		<div class="field fw5">Go back to step 6 (repeat 15x)</div>
		<div class="field fw2"></div>
                <div class="field fw2"></div>
	</div>
	<div class="record">
		<div class="field fw1">10</div>
		<div class="field fw5">Final Extention</div>
		<div class="field fw2">72 °C</div>
                <div class="field fw2">10:00</div>
	</div>
	</div>
	<div class="caption">Recommended thermocycle.</div>
</div>
</html>
}}

{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 19 May 2015 - 00:00, 19 May 2015
	|executor=Harm Ruesink
	|content=<html>
<div class="text"> Ran a gel with PCR 3 products on 1% agarose gel with EtBr. 
Both sample 1 and sample 2 show no bands visible. Probably because yuaB forward 
primer doesn’t work. </div>
</html>
}}


{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 21 May 2015 - 00:00, 21 May 2015
	|executor=Harm Ruesink
	|content=<html>
<div class="text"> Ordered new primers. </div>
<div class="object data" id="tbl1">
	<div class="wrapper">
	<div class="header">
		<div class="field fw3">Primer</div>
		<div class="field fwx">Sequence</div>
	</div>
	<div class="record">
		<div class="field fw3">yuaB forward 2 (with RBS)</div>
		<div class="field 
fwx">GCGAATTCGCGGCCGCTTCTAGAGTTAGGGGGAATTTTGTTATGAAACGC</div>
	</div>
	<div class="record">
		<div class="field fw3">yuaB forward 3 (no RBS)</div>
		<div class="field 
fwx">GCGAATTCGCGGCCGCTTCTAGAGATGAAACGCAAATTATTATCTTCTTTGG</div>
	</div>
	</div>
	<div class="caption">New primers.</div>
</div>
</html>
}}

{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 8 June 2015 - 00:00, 8 June 2015
	|executor=Harm Ruesink
	|content=<html>
<div class="subtitle">PCR 5</div>
<div class="text">PCR'd part 1 of yuaB using primers yuaB forward 2 & yuaB 
forward 3 and genomic DNA from B. subtilis 168.</div>
<div class="object data" id="tbl1">
	<div class="wrapper">
	<div class="header">
		<div class="field fw3">Ingredient</div>
		<div class="field fw3">Quantity</div>
	</div>
	<div class="record">
		<div class="field fw3">10x buffer</div>
		<div class="field fw3">10 µL</div>
	</div>
	<div class="record">
		<div class="field fw3">dNTP MM (2mM)</div>
		<div class="field fw3">10 µL</div>
	</div>
	<div class="record">
		<div class="field fw3">Dreamtaq polymerase</div>
		<div class="field fw3">1 µL</div>
	</div>
	<div class="record">
		<div class="field fw3">\( \mathrm{H_2 O} \)</div>
		<div class="field fw3">76.5 µL</div>
	</div>
	</div>
	<div class="caption">PCR 5</div>
</div>
<div class="text">
The following mix was used for each sample.
</div>
<div class="object data" id="tbl1">
	<div class="wrapper">
	<div class="header">
		<div class="field fw3">Ingredient</div>
		<div class="field fw3">Quantity</div>
	</div>
	<div class="record">
		<div class="field fw3">Mastermix PCR 5</div>
		<div class="field fw3">0,3 µL per template DNA</div>
	</div>
	<div class="record">
		<div class="field fw3">Per primer</div>
		<div class="field fw3">0,3 µL</div>
	</div>
	</div>
	<div class="caption">Sample contents.</div>
</div>
<div class="text">
With this mix the following samples were made:
</div>
<div class="listing">
<div class="item">2. primers: yuaB forward 2 + yuaB overlap rev, template DNA: 
B. subtilis 168 DNA</div>
<div class="item">3. primers: yuaB forward 3 + yuaB overlap rev, template DNA: 
B. subtilis 168 DNA</div>    
</div>
<div class="text">
The following PCR thermocycle was performed.
</div>
<div class="object data" id="tbl1">
	<div class="wrapper">
	<div class="header">
		<div class="field fw1">#</div>
		<div class="field fw5">Step</div>
		<div class="field fw2">Temperature</div>
                <div class="field fw2">Time</div>
	</div>
	<div class="record">
		<div class="field fw1">7</div>
		<div class="field fw5">Initial Denaturation</div>
		<div class="field fw2">98 °C</div>
                <div class="field fw2">3:00</div>
	</div>
	<div class="record">
		<div class="field fw1">8</div>
		<div class="field fw5">Denaturation</div>
		<div class="field fw2">98 °C</div>
                <div class="field fw2">0:30</div>
	</div>
	<div class="record">
		<div class="field fw1">9</div>
		<div class="field fw5">Annealing</div>
		<div class="field fw2">60 °C</div>
                <div class="field fw2">0:30</div>
	</div>
	<div class="record">
		<div class="field fw1">10</div>
		<div class="field fw5">Extention</div>
		<div class="field fw2">72 °C</div>
                <div class="field fw2">1:00</div>
	</div>
	<div class="record">
		<div class="field fw1">11</div>
		<div class="field fw5">Go back to step 2 (repeat 30x)</div>
		<div class="field fw2"></div>
                <div class="field fw2"></div>
	</div>
	<div class="record">
		<div class="field fw1">12</div>
		<div class="field fw5">Final Extension</div>
		<div class="field fw2">72 °C</div>
                <div class="field fw2">10:00</div>
	</div>	
    </div>
    <div class="caption">Recommended thermocycle.</div>
</div>
<div class="text">
Loaded PCR product on 1% agarose gel with DNA stain G (1:50000) and ran for 1 
hour on 100V. Also loaded sample 1.2  (yuaB part 2 of length 209 bp). 
The following samples were prepared.
</div>
<div class="listing">
    <div class="item">
        30 µL DNA(tasA) 
    </div>
    <div class="item">
        6 µL 6x buffer
    </div>
</div>
<div class="text">
10µL of Quick-Load® Purple 2-Log DNA Ladder (0.1 - 10.0 kb) was used. 
</div>
<div class="object data" id="tbl2">
    <div class="wrapper">
    <div class="header">
        <div class="field fw3">Sample</div>
        <div class="field fwx">Result</div>
    </div>
    <div class="record">
        <div class="field fw3">2</div>
        <div class="field fwx">bands visible of ~450 bp</div>
    </div>
    <div class="record">
        <div class="field fw3">3</div>
        <div class="field fwx">bands visible of ~450 bp</div>
    </div>
    <div class="record">
        <div class="field fw3">1.2</div>
        <div class="field fwx">bands visible of ~200 bp</div>
    </div>
    </div>
    <div class="caption">PCR results</div>
</div>
<div class="text">
Bands were cut from the gel and purified using a PCR purification kit. 
</div>
<div class="object data" id="tbl3">
    <div class="wrapper">
    <div class="header">
        <div class="field fw3">Sample</div>
        <div class="field fw3">Gel Weight</div>
        <div class="field fw3">Binding buffer</div>
    </div>
    <div class="record">
        <div class="field fw3">2 (1.2)</div>
        <div class="field fw3">253 mg</div>
        <div class="field fw3">506 µL</div>
    </div>
    <div class="record">
        <div class="field fw3">3 (1.3)</div>
        <div class="field fw3">193 mg</div>
        <div class="field fw3">386 µL</div>
    </div>
    <div class="record">
        <div class="field fw3">1.2 (2)</div>
        <div class="field fw3">270 mg</div>
        <div class="field fw3">540 µL</div>
    </div>
</div>
    <div class="caption">Gels.</div>
</div>
<div class="text">
Samples were stored as
</div>
<div class="listing"> 
    <div class="item">1.2 (49,9 ng/µL Nanodrop value)</div>
    <div class="item">1.3 (56,5 ng/µL Nanodrop value)</div>
    <div class="item">2 (19,2 ng/µL Nanodrop value)</div>
</div>
</html>
}}

{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 9 June 2015 - 00:00, 9 June 2015
	|executor=Harm Ruesink
	|content=<html>
<div class="subtitle">Assembly PCR</div>
<div class="text">Part 1 (450 bp) and part 2 (209 bp) were combined to get the 
whole bslA gene.</div>
<div class="object data" id="tbl4">
    <div class="wrapper">
    <div class="header">
        <div class="field fw5">Ingredient</div>
        <div class="field fw2">Quantity</div>
    </div>
    <div class="record">
        <div class="field fw5">10x buffer</div>
        <div class="field fw2">10 µL</div>
    </div>
    <div class="record">
        <div class="field fw5">dNTP MM (2 mM)</div>
        <div class="field fw2">10 µL</div>
    </div>
    <div class="record">
        <div class="field fw5">Dreamtaq polymerase</div>
        <div class="field fw2">1 µL</div>
    </div>
    <div class="record">
        <div class="field fw5">\( \mathrm{H_2 O}\)</div>
        <div class="field fw2">78 µL</div>
    </div>
    </div>
    <div class="caption">PCR Mastermix.</div>
</div>
<div class="text">
Per sample the following mix was made.
</div>
<div class="listing">
    <div class="item">30 µL of mastermix</div>
    <div class="item">1 µL per template DNA</div>
    <div class="item">0.3 µL per primer</div>
</div>
<div class="text">
Using this mix two samples were prepared.
</div>
<div class="object data" id="tbl5">
    <div class="wrapper">
    <div class="header">
        <div class="field fw6">Primers</div>
        <div class="field fw3">Template DNA</div>
    </div>
    <div class="record">
        <div class="field fw6">yuab forward 2 + yuaB rev</div>
        <div class="field fw3">1.3 + 2</div>
    </div>
    <div class="record">
        <div class="field fw6">yuaB forward 3 + yuaB rev</div>
        <div class="field fw3">1.3 + 2</div>
    </div>
    </div>
    <div class="caption">Samples.</div>
</div>

<div class="object data" id="tbl6">
    <div class="wrapper">
    <div class="header">
        <div class="field fw1">#</div>
        <div class="field fw5">Step</div>
        <div class="field fw5">Temperature</div>
        <div class="field fw2">Time</div>
    </div>
    <div class="record">
        <div class="field fw1">1</div>
        <div class="field fw5">Denaturation</div>
        <div class="field fw5">98 °C</div>
        <div class="field fw2">3:00</div>
    </div>
    <div class="record">
        <div class="field fw1">2</div>
        <div class="field fw5">Denaturation</div>
        <div class="field fw5">98 °C</div>
        <div class="field fw2">0:30</div>
    </div>
    <div class="record">
        <div class="field fw1">3</div>
        <div class="field fw5">Annaeling</div>
        <div class="field fw5">60 °C</div>
        <div class="field fw2">0:30</div>
    </div>
    <div class="record">
        <div class="field fw1">4</div>
        <div class="field fw5">Extention</div>
        <div class="field fw5">72 °C</div>
        <div class="field fw2">1:00</div>
    </div>
    <div class="record">
        <div class="field fw1">5</div>
        <div class="field fw5">Go back to step 2 (repeat 30x)</div>
        <div class="field fw5"></div>
        <div class="field fw2"></div>
    </div>
    <div class="record">
        <div class="field fw1">6</div>
        <div class="field fw5">Final Extension</div>
        <div class="field fw5">72 °C</div>
        <div class="field fw2">10:00</div>
    </div>
    </div>
    <div class="caption">Recommended thermocycle.</div>
</div>
<div class="text">PCR product was loaded on 1% agarose gel with DNA stain G 
(1:50000) and ran for one hour at 100 V. The following samples were 
analyzed.</div>
<div class="listing">
    <div class="item">30 µL of DNA (tasA)</div>
    <div class="item">6 µL of 6x buffer</div>
</div>
<div class="text">
10 µL of Quick-Load® Purple 2-Log DNA Ladder (0.1 - 10.0 kb) was used. Sample 1 
showed bands at ~ 650 bp. Sample 2 showed bands at ~650 bp. Band were cut from 
the gel and purified using a PCR purification kit.</div>
<div class="object data" id="tbl7">
    <div class="wrapper">
    <div class="header">
        <div class="field fw2">Sample</div>
        <div class="field fw3">Gel Weight</div>
        <div class="field fw3">Binding Buffer</div>
    </div>
    <div class="record">
        <div class="field fw2">1</div>
        <div class="field fw3">176 mg</div>
        <div class="field fw3">352 µL</div>
    </div>
    <div class="record">
        <div class="field fw2">2</div>
        <div class="field fw3">248 mg</div>
        <div class="field fw3">496 µL</div>
    </div>
    </div>
    <div class="caption">Purification.</div>
</div>
<div class="text"> This was stored (20 µL of elution buffer) as follows.</div>
<div class="listing">
    <div class="item">bslA + RBS, 9-6-15</div>
    <div class="item">bsLA - RBS, 96-15</div>
</div>
</html>
}}

{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 10 June 2015 - 00:00, 10 June 2015
	|executor=Harm Ruesink
	|content=<html>
<div class="subtitle">Digestion</div>
<div class="text">yuaB was digested using the following mix.</div>
<div class="object data" id="tbl8">
    <div class="wrapper">
    <div class="header">
        <div class="field fw5">Ingredient</div>
        <div class="field fw2">Quantity</div>
    </div>
    <div class="record">
        <div class="field fw5">DNA (yuaB + RBS)</div>
        <div class="field fw2">2 µL</div>
    </div>
    <div class="record">
        <div class="field fw5">10x buffer (2.1)</div>
        <div class="field fw2">2 µL</div>
    </div>
    <div class="record">
        <div class="field fw5">EcoRI</div>
        <div class="field fw2">1 µL</div>
    </div>
    <div class="record">
        <div class="field fw5">Pstl</div>
        <div class="field fw2">1 µL</div>
    </div>
    <div class="record">
        <div class="field fw5">\( \mathrm{H_2 O} \)</div>
        <div class="field fw2">14 µL</div>
    </div>
    </div>
    <div class="caption">Digestion mix.</div>
</div>
<div class="text">
The mix was incubated for two hours at 37 °C. A 1% agarose + DNA strain G 
1:50000 was ran at 120 V. </div>
<div class="object data" id="tbl9">
    <div class="wrapper">
    <div class="header">
        <div class="field fw3">Ingredient</div>
        <div class="field fw2">Quantity</div>
    </div>
    <div class="record">
        <div class="field fw3">DNA</div>
        <div class="field fw2">20 µL</div>
    </div>
    <div class="record">
        <div class="field fw3">6x buffer</div>
        <div class="field fw2">4 µL</div>
    </div>
    </div>
    <div class="caption">Sample.</div>
</div>
<div class="text">
10 µL of Quick-Load® Purple 2-Log DNA Ladder (0.1 - 10.0 kb) was used. A band 
was visible around 600 bp. This means that the restriction site within the yuaB 
gene is gone. </div>
<div class="text">
DNA was extracted from the gel using a PCR purification kit.
</div>
<div class="object data" id="tbl7">
    <div class="wrapper">
    <div class="header">
        <div class="field fw2">Sample</div>
        <div class="field fw3">Gel Weight</div>
        <div class="field fw3">Binding Buffer</div>
    </div>
    <div class="record">
        <div class="field fw2">yuaB</div>
        <div class="field fw3">225 mg</div>
        <div class="field fw3">450 µL</div>
    </div>
    </div>
    <div class="caption">Purification.</div>
</div>
<div class="text">Eluted in 10 µL elution buffer and stored as "yuaB dig". 
</div>
<div class="text">
Ligation of yuaB in BBa_K823023 was performed using the following ligation 
mix.</div>
<div class="object data" id="tbl8">
    <div class="wrapper">
    <div class="header">
        <div class="field fw5">Ingredient</div>
        <div class="field fw1">Quantity</div>
    </div>
    <div class="record">
        <div class="field fw5">insert DNA (yuaB dig)</div>
        <div class="field fw1">10 µL</div>
    </div>
    <div class="record">
        <div class="field fw5">vector DNA (BBa_K823023)</div>
        <div class="field fw1">5 µL</div>
    </div>
    <div class="record">
        <div class="field fw5">10x buffer</div>
        <div class="field fw1">2 µL</div>
    </div>
    <div class="record">
        <div class="field fw5">T4 ligase</div>
        <div class="field fw1">0.5 µL</div>
    </div>
    <div class="record">
        <div class="field fw5">\(\mathrm{H_2 O}\)</div>
        <div class="field fw1">2.5 µL</div>
    </div>
    </div>
    <div class="caption">Ligation mix</div>
</div>
<div class="text">This was ligated for 2 hours at room temperature. The DNA was 
then transformed into competent E. coli using heat shock transformation. 2 
hours 
later it was plated on LB ampicilin plates and left in 37 °C overnight. </div>
</html>
}}

{{Team:Groningen/TEMPLATES/LOG
	|title=16:00, 11 June 2015 - 16:30, 11 June 2015
	|executor=Harm Ruesink
	|content=<html>
<div class="text">Two white colonies and a lot of red colonies visible on 
plate. 
Inoculated 2 white colonies and one red colony in liquid LB with ampicilin 
overnight.</div>
</html>
}}

{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 12 June 2015 - 00:00, 12 June 2015
	|executor=Harm Ruesink
	|content=<html>
<div class="text">Miniprepped overnight cultures. Stored as </div>
<div class="listing">
    <div class="item">
    23 yuaB 1
    </div>
    <div class="item">
    23 yuab 2
    </div>
    <div class="item">
    23 RFP
    </div>
</div>
</html>
}}

{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 15 June 2015 - 00:00, 15 June 2015
	|executor=Harm Ruesink
	|content=<html>
<div class="text">Digested yuaB from backbone to check ligation using EcoRI and 
Pstl. The following mastermix was prepared for 13 samples.</div>
<div class="object data" id="tbl8">
    <div class="wrapper">
    <div class="header">
        <div class="field fw5">
        Ingredient
        </div>
        <div class="field fw3">
        Quantity
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
        10x buffer (2.1)
        </div>
        <div class="field fw3">
        30 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
        EcorI
        </div>
        <div class="field fw3">
        10 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
        Pslt
        </div>
        <div class="field fw3">
        10 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
        \(\mathrm{H_2 O}\)
        </div>
        <div class="field fw3">
        185 µL
        </div>
    </div>
    </div>
    <div class="caption">Digestion mastermix.</div>
</div>
<div class="text">
Per sample the following mix was prepared.
</div>
<div class="object data" id="tbl9">
    <div class="wrapper">
    <div class="header">
        <div class="field fw5">
        Ingredient
        </div>
        <div class="field fw3">
        Quantity
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
        DNA
        </div>
        <div class="field fw3">
        15 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
        Mastermix
        </div>
        <div class="field fw3">
        15 µL
        </div>
    </div>
    </div>
    <div class="caption">Sample contents.</div>
</div>
<div class="text">The samples were incubated at 37 °C for two hours. They were 
then loaded on 1% agarose gel with 1:50000 DNA stain G. To 20 µL of digestion 
sample 4 µL of 6x buffer was added. 10µL of Quick-Load® Purple 2-Log DNA Ladder 
(0.1 - 10.0 kb) was used for each sample.   </div>
<div class="text">
Nothing was seen on the gel. The gel may have been too hot when adding the DNA 
stain G.
</div>
</html>
}}

{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 16 June 2015 - 00:00, 16 June 2015
	|executor=Harm Ruesink
	|content=<html>
<div class="text"> Repeated digest of tasA from 15-6-15. Digest for 1.5h at 37 
°C.
</div>
<div class="text">10 µL of digestion sample was added to 2 L of 6x buffer. This 
was run at 100 V for 1 hour with 10 µL of 2log ladder. </div>
<div class="text">
No insert was seen on gel. yuaB probably wasn't ligated in the backbone.
</div>
</html>
}}
{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 8 July 2015 - 00:00, 8 July 2015
	|executor=Harm Ruesink
	|content=<html>
<div class="subtitle">Ligation of yuaB into pSB1C3</div>
<div class="text">yuaB + RBS with pSB1C3 was digested using EcoRI and Pstl. 
</div>
<div class="object data" id="tbl10">
    <div class="wrapper">
    <div class="header">
        <div class="field fw5">
        Ingredient
        </div>
        <div class="field fw3">
        Quantity
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
        10x buffer (2.1)
        </div>
        <div class="field fw3">
        6.6 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
        EcoRI
        </div>
        <div class="field fw3">
        3.3 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
        Pstl
        </div>
        <div class="field fw3">
        3.3 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
        \(\mathrm{H_2 O}\)
        </div>
        <div class="field fw3">
        23.4 µL
        </div>
    </div>
    </div>
    <div class="caption">Ligation mastermix.</div>
</div>
<div class="object data" id="tbl11">
    <div class="wrapper">
    <div class="header">
        <div class="field fw1">
        Sample
        </div>
        <div class="field fw3">
        Ingredient
        </div>
        <div class="field fw3">
        Quantity
        </div>
    </div>
    <div class="record">
        <div class="field fw1">
        1
        </div>
        <div class="field fw3">
        yuaB + RBS
        </div>
        <div class="field fw3">
        10 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw1">
        
        </div>
        <div class="field fw3">
        Mastermix
        </div>
        <div class="field fw1">
        10 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw1">
        2
        </div>
        <div class="field fw3">
        pSB1C3
        </div>
        <div class="field fw3">
        10 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw1">
        
        </div>
        <div class="field fw3">
        Mastermix
        </div>
        <div class="field fw1">
        10 µL
        </div>
    </div>
    </div>
    <div class="caption">Samples.</div>
</div>
<div class="text">This was digested for 1 h at 37 °C. The DNA was then purified 
using a PCR purification kit. yuaB was then ligated in pSB1C3. </div>

<div class="object data">
    <div class="wrapper">
    <div class="header">
        <div class="field fw1">
        Sample
        </div>
        <div class="field fw5">
        Ingredient
        </div>
        <div class="field fw3">
        Quantity
        </div>
    </div>
    <div class="record">
        <div class="field fw1">
        2
        </div>
        <div class="field fw5">
        bslA DNA
        </div>
        <div class="field fw3">
        10 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw1">
        
        </div>
        <div class="field fw5">
        pSB1C3 DNA
        </div>
        <div class="field fw3">
        5 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw1">
        
        </div>
        <div class="field fw5">
        T4 ligase
        </div>
        <div class="field fw3">
        0.5 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw1">
        
        </div>
        <div class="field fw5">
        T4 ligase buffer
        </div>
        <div class="field fw3">
        2 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw1">
        
        </div>
        <div class="field fw5">
        \(\mathrm{H_2 O}\)
        </div>
        <div class="field fw3">
        2.5 µL
        </div>
    </div>
    </div>
    <div class="caption">Sample.</div>
</div>
<div class="text">The sample was ligated at room temperature for two hours. The 
ligation product was then transformed into 100 µL of NED competent cells using 
heat shock transformation. Two plates were made.</div>
<div class="listing">
    <div class="item">
    90% plate (90 µL)
    </div>
    <div class="item">
    10% plate (10 µL)
    </div>
</div>
<div class="text">
    Plates were incubated overnight at 37 °C.
</div>
</html>
}}

{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 9 Juli 2015 - 00:00, 9 Juli 2015
	|executor=Harm Ruesink
	|content=<html>
<div class="subtitle">Transformation results</div>
<div class="text">Transformation was a success, both plates show a large number 
of colonies. Four colonies from the 10% plate were grown overnight in 3 mL LB 
with 6 µL of 1/500 chloramphenicol</div>
</html>
}}
{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 16 July 2015 - 00:00, 16 July 2015
	|executor=Harm Ruesink
	|content=<html>
<div class="text">Ligation of a salt promotor and yuaB into BBa_K823023 was 
performed. </div>
</html>
<div class="object data">
    <div class="wrapper">
    <div class="header">
        <div class="field fw1">
            Sample
        </div>
        <div class="field fw5">
            DNA
        </div>
        <div class="field fw3">
            Restriction enzyme
        </div>
    </div>
    <div class="record">
        <div class="field fw1">
            1
        </div>
        <div class="field fw5">
            Psalt 
        </div>
        <div class="field fw3">
            EcoRI + Spel
        </div>
    </div>
    <div class="record">
        <div class="field fw1">
            2
        </div>
        <div class="field fw5">
            bslA (y4, 10-7-15)
        </div>
        <div class="field fw3">
            Xbal + PstI
        </div>
    </div>
    <div class="record">
        <div class="field fw1">
            3
        </div>
        <div class="field fw5">
            BBa_K823023
        </div>
        <div class="field fw3">
            EcoRI + PstI
        </div>
    </div>
    </div>
    <div class="caption">Digest.</div>
</div>

<div class="object data">
    <div class="wrapper">
    <div class="header">
        <div class="field fw2">
            Sample
        </div>
        <div class="field fw5">
            Ingredient
        </div>
        <div class="field fw2">
            Quantity
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            1
        </div>
        <div class="field fw5">
            DNA (Psalt)
        </div>
        <div class="field fw2">
            6 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw5">
            EcoRI
        </div>
        <div class="field fw2">
            1 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw5">
            Spel
        </div>
        <div class="field fw2">
            1 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw5">
            10x buffer (2.1)
        </div>
        <div class="field fw2">
            2 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw5">
            \(\mathrm{H_2 O}\)
        </div>
        <div class="field fw2">
            10 µL
        </div>
    </div>
        
    <div class="record">
        <div class="field fw2">
            2
        </div>
        <div class="field fw5">
            DNA (Y4)
        </div>
        <div class="field fw2">
            3 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw5">
            Xbal
        </div>
        <div class="field fw2">
            1 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw5">
            PstI
        </div>
        <div class="field fw2">
            1 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw5">
            10x buffer (2.1)
        </div>
        <div class="field fw2">
            2 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw5">
            \(\mathrm{H_2 O}\)
        </div>
        <div class="field fw2">
            13 µL
        </div>
    </div>

    <div class="record">
        <div class="field fw2">
            2
        </div>
        <div class="field fw5">
            DNA (K823023)
        </div>
        <div class="field fw2">
            3 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw5">
            EcoRI
        </div>
        <div class="field fw2">
            1 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw5">
            PstI
        </div>
        <div class="field fw2">
            1 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw5">
            10x buffer (2.1)
        </div>
        <div class="field fw2">
            2 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw5">
            \(\mathrm{H_2 O}\)
        </div>
        <div class="field fw2">
            10 µL
        </div>
    </div>
    </div>
    <div class="caption">
        Samples.
    </div>
</div>
<div class="text">Samples were incubated for one hour at 37 °C. They were then 
purified using a PCR purification kit (20 µL elution). Samples were stored as 
"1 
& 2" with "16-7-15" on the side.</div>

<div class="text"> Samples 1 + 2 + 3 were ligated. </div>
<div class="object data">
    <div class="wrapper">
    <div class="header">
        <div class="field fw5">
            Ingredient
        </div>
        <div class="field fw3">
            Quantity
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
            DNA 1
        </div>
        <div class="field fw3">
            5 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
            DNA 2
        </div>
        <div class="field fw3">
            5 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
            T4 Ligase
        </div>
        <div class="field fw3">
            0.5 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
            10x buffer
        </div>
        <div class="field fw3">
            2 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
            \(\mathrm{H_2 O}\)
        </div>
        <div class="field fw3">
            5 µL
        </div>
    </div>
    </div>
    <div class="caption">Ligation mix.</div>
</div>

<div class="text">Ligated at room temperature for 30 minutes. Transformed into 
10 µL of NEB competent cells with 10 µL of ligation product using heat shock 
transformation. The result was plated on LB ampicilin plates and left at 37 °C 
overnight.</div>
}}



{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 17 July 2015 - 00:00, 17 July 2015
	|executor=Harm Ruesink
	|content=<html>
<div class="subtitle">Transformation results</div>
<div class="text"> A few white colonies were seen on the plate. A colony PCR 
was 
performed to check the size of the insert (which is supposed to be bslA + 
Psalt). The colony PCR showed possible bslA + Psalt insert in one sample. 
Colony 
was grown overnight in LB + ampicilin.
</div>
</html>
}}


{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 18 August 2015 - 00:00, 18 August 2015
	|executor=Harm Ruesink
	|content=<html>
	<div class="text">
            Miniprep was performed and stored as "22 (17-7-15)".    
        </div>
</html>
}}



{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 20 July 2015 - 00:00, 20 July 2015
	|executor=Harm Ruesink
	|content=<html>
<div class="text">Send in sample 22 for sequencing with prefix (19) and suffix 
(20) primers. </div>
<div class="text">Send sample 22 to Macrogen for sequencing.</div>
<div class="object data">
    <div class="wrapper">
    <div class="header">
        <div class="field fw5">
            Ingredient
        </div>
        <div class="field fw2">
            Quantity
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
            1/20 primer (5 pM/µL)
        </div>
        <div class="field fw2">
            5 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
            plasmid DNA (22)
        </div>
        <div class="field fw2">
            5 µL
        </div>
    </div>
    </div>
    <div class="caption">Samples.</div>
</div>
<div class="listing">
    <div class="item">
        Forward primer (prefix primer): 17C4ZAD516
    </div>
    <div class="item">
        Reverse primer (suffix primer): 17C4ZAD517
    </div>
</div>
</html>
}}



{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 21 July 2015 - 00:00, 21 July 2015
	|executor=Harm Ruesink
	|content=<hmtl>
<div class="text">Transformed sample 22 into B. subtilis.	 Took B. 
subtilis 3610 ComI colony from plate (13-7-15 in fridge) and put it in MC 1x 
for 
5 hours. Optical density at 600 nm was measured to be 0.15. Then 5 µL of 
plasmid 
DNA (22) was added to 400 µL of cell culture and left in 37 °C
 shaking incubator for a further two hours. The culture was then plated on LB 
chloramphenicol plates and left overnight at 37 °C.
</div>
</html>
}}
{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 22 July 2015 - 00:00, 22 July 2015
	|executor=Harm Ruesink
	|content=<html>
<div class="subtitle">Sequencing results</div>
<div class="text">Results don't show correct insert. Insert is an RFP. 
Discarded 
transformed Bacillus subtilis due to incorrect insert. </div>
</html>
}}



{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 23 July 2015 - 00:00, 23 July 2015
	|executor=Harm Ruesink
	|content=<html>
<div class="text"> Psalt was digsted at for one huur at 37 °C using the 
following mix.</div>
<div class="object data">
    <div class="wrapper">
    <div class="header">
        <div class="field fw4">
            Ingredient
        </div>
        <div class="field fw2">
            Quantity
        </div>
    </div>
    <div class="record">
        <div class="field fw4">
            DNA (Psalt)
        </div>
        <div class="field fw2">
            10 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw4">
            EcoRI
        </div>
        <div class="field fw2">
            1 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw4">
            Spel
        </div>
        <div class="field fw2">
            1 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw4">
            10x buffer (2.1)
        </div>
        <div class="field fw2">
            2 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw4">
            \(\mathrm{H_2 O}\)
        </div>
        <div class="field fw2">
            6 µL
        </div>
    </div>
    </div>
    <div class="caption">Digestion mix</div>
</div>
<div class="text">The digestion sample was purified using a PCR purification 
kit. Eluted with 20 µL. It was then ligated for two hours at room temperature 
using the following mix. </div> <div class="object data">
    <div class="wrapper">
    <div class="header">
        <div class="field fw5">
            Ingredient
        </div>
        <div class="field fw2">
            Quantity
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
            DNA 1 (Psalt)
        </div>
        <div class="field fw2">
            5 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
            DNA 2 (bslA)
        </div>
        <div class="field fw2">
            5 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
            DNA 3 (K23)
        </div>
        <div class="field fw2">
            5 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
            T4 Ligase
        </div>
        <div class="field fw2">
            1 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
            10x buffer
        </div>
        <div class="field fw2">
            2 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
            \(\mathrm{H_2 O}\)
        </div>
        <div class="field fw2">
            2 µL
        </div>
    </div>
    </div>
    <div class="caption">Ligation mix.</div>
</div>
<div class="text">A heat shock transformation was then performed. The result 
was 
plated on LB ampicilin and incubated overnight at 37 °C. </div>
</html>
}}


{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 24 July 2015 - 00:00, 24 July 2015
	|executor=Harm Ruesink
	|content=<html>
	<div class="subtitle">Transformation Results</div>
	<div class="text">
	No white colonies were visible on the plate.
	</div>
</html>
}}

{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 28 July 2015 - 00:00, 28 July 2015
	|executor=Harm Ruesink
	|content=<html>
        <div class="subtitle">Adding promoters to Y4 (10-7-15).</div>
        <div class="listing">
            <div class="item">
                Pveg (iGEM distribution kit 2015, plate 1, 20 G)
            </div>
        </div>
        <div class="text"> For the promoters the restriction enzymes EcoRI & 
Spel were used. For the backbone Xbal & EcoRI were used. 
        </div>
    <div class="object data">
        <div class="wrapper">
        <div class="header">
            <div class="field fw2">
                Sample
            </div>
            <div class="field fw5">
                Ingredient
            </div>
            <div class="field fw2">
                Quantity
            </div>
        </div>
        <div class="record">
            <div class="field fw2">
                1
            </div>
            <div class="field fw5">
                Y4 DNA
            </div>
            <div class="field fw2">
                5 µL
            </div>
        </div>
        <div class="record">
            <div class="field fw2">
                
            </div>
            <div class="field fw5">
                10x buffer
            </div>
            <div class="field fw2">
                2 µL
            </div>
        </div>
        <div class="record">
            <div class="field fw2">
                
            </div>
            <div class="field fw5">
               EcoRI
            </div>
            <div class="field fw2">
                1 µL
            </div>
        </div>
        <div class="record">
            <div class="field fw2">
                
            </div>
            <div class="field fw5">
                Xbal
            </div>
            <div class="field fw2">
                1 µL
            </div>
        </div>
        <div class="record">
            <div class="field fw2">
                
            </div>
            <div class="field fw5">
                \(\mathrm{H_2 O}\)
            </div>
            <div class="field fw2">
                11 µL
            </div>
        </div>
        <div class="record">
            <div class="field fw2">
                3
            </div>
            <div class="field fw5">
                Pveg DNA
            </div>
            <div class="field fw2">
                10 µL
            </div>
        </div>
        <div class="record">
            <div class="field fw2">
                
            </div>
            <div class="field fw5">
                10x buffer
            </div>
            <div class="field fw2">
                2 µL
            </div>
        </div>
        <div class="record">
            <div class="field fw2">
                
            </div>
            <div class="field fw5">
                EcoRI
            </div>
            <div class="field fw2">
                1 µL
            </div>
        </div>
        <div class="record">
            <div class="field fw2">
                
            </div>
            <div class="field fw5">
                Spel
            </div>
            <div class="field fw2">
                1 µL
            </div>
        </div>
        <div class="record">
            <div class="field fw2">
                
            </div>
            <div class="field fw5">
                \(\mathrm{H_2 O}\)
            </div>
            <div class="field fw2">
                11 µL
            </div>
        </div>
        </div>
        <div class="caption">Digestion samples</div>
</div>
        <div class="text">These samples were digested at 37 °C for two hours. 
They were then on a gel at 100 V for 45 minutes. 1% agarose with DNA stain G 
1:50000 was used. For each sample, 20 µL of DNA was mixed with 4 µL of 6x 
buffer. 10 µL of Quick-Load® Purple 2-Log DNA Ladder (0.1 - 10.0 kb) was used. 
</div> 
<div class="object data">
    <div class="wrapper">
    <div class="header">
        <div class="field fw2">
            Sample
        </div>
        <div class="field fw4">
            Weight
        </div>
        <div class="field fw4">
            Binding buffer
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            1
        </div>
        <div class="field fw4">
            156 mg
        </div>
        <div class="field fw4">
            316 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            3
        </div>
        <div class="field fw4">
            223 mg
        </div>
        <div class="field fw4">
            446 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            4
        </div>
        <div class="field fw4">
            300 mg
        </div>
        <div class="field fw4">
            600 µL
        </div>
    </div>
    </div>
    <div class="caption">Gel cuts.</div>
</div>
<div class="text">
    The DNA was purified using a PCR purification kit (Genejet #K0701, lot 
00265976). 25 µL elution buffer was used. The concentrations were checked using 
Nanodrop.
</div>
<div class="object data">
    <div class="wrapper">
    <div class="header">
        <div class="field fw3">
            Sample
        </div>
        <div class="field fw3">
            Concentration
        </div>
    </div>
    <div class="record">
        <div class="field fw3">
            Y4 dig (1)
        </div>
        <div class="field fw3">
            2 ng/µL
        </div>
    </div>
    <div class="record">
        <div class="field fw3">
            Pveg (3)
        </div>
        <div class="field fw3">
            1.4 ng/µL
        </div>
    </div>
    <div class="record">
        <div class="field fw3">
            Psalt (4)
        </div>
        <div class="field fw3">
            5 ng/µL
        </div>
    </div>
    </div>
    <div class="caption">Sample concentrations.</div>
</div>

<div class="subtitle">Ligation of promoters in backbones</div>

<div class="object data">
    <div class="wrapper">
    <div class="header">
        <div class="field fw4">
            Ingredient
        </div>
        <div class="field fw2">
            Quantity
        </div>
    </div>
    <div class="record">
        <div class="field fw4">
            10x buffer
        </div>
        <div class="field fw2">
            12 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw4">
            T4 Ligase
        </div>
        <div class="field fw2">
            4 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw4">
            \(\mathrm{H_2 O}\)
        </div>
        <div class="field fw2">
            8 µL
        </div>
    </div>
    </div>
    <div class="caption">Ligation mastermix.</div>
</div>
<div class="object data">
    <div class="wrapper">
    <div class="header">
        <div class="field fw2">
            Sample
        </div>
        <div class="field fw4">
            Ingredient
        </div>
        <div class="field fw2">
            Quantity
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            1
        </div>
        <div class="field fw4">
            Y4 dig (1)
        </div>
        <div class="field fw2">
            12 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw4">
            Pveg (3)
        </div>
        <div class="field fw2">
            12 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw4">
            Mastermix
        </div>
        <div class="field fw2">
            6 µL
        </div>
    </div>
    
    <div class="record">
        <div class="field fw2">
            2
        </div>
        <div class="field fw4">
            Y4 dig (1)
        </div>
        <div class="field fw2">
            12 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw4">
            Psalt (4)
        </div>
        <div class="field fw2">
            12 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw4">
            Mastermix
        </div>
        <div class="field fw2">
            6 µL
        </div>
    </div>
    </div>
    <div class="caption">Ligation samples.</div>
</div>
<div class="text">Samples were ligated overnight.</div>
</html>
}}


{{Team:Groningen/TEMPLATES/LOG
	|title=10:00, 29 July 2015 - 12:00, 29 July 2015
	|executor=Harm Ruesink
	|content=<html>
<div class="text">Heat shock transformation was performed. Samples were plated 
on LB chloramphenicol and incubated overnight at 37 °C. </div>
</html>
}}



{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 30 July 2015 - 00:00, 30 July 2015
	|executor=Harm Ruesink
	|content=<html>
<div class="subtitle">Transformation results</div>
<div class="text">No colonies visible on LB chloramphenicol plate.</div>
<div class="text">Y4 was send to Macrogen for sequencing.</div>
<div class="listing">
    <div class="item">5 µL of 1/20 primer (5 pmole/µL)</div>
    <div class="item">5 µL of plasmid DNA (Y4) </div>
</div>
<div class="subtitle">Sequencing codes Macrogen</div>
<div class="listing">
    <div class="item">
    Forward primer (prefix (19)): 17C4ZAD660
    </div>
    <div class="item">
    Reverse primer (suffix (20)): 17C4ZAD661
    </div>
</div>
</html>
}}

{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 6 August 2015 - 00:00, 6 August 2015
	|executor=Harm Ruesink
	|content=<html>
<div class="text">Digestion and ligation of Pliag in front of Y4 was performed. 
For the promoters EcoRI and Spel were used. For Y4 Xbal and EcoRI were 
used. These enzymes leave a scar.</div>
<div class="object data">
    <div class="wrapper">
    <div class="header">
        <div class="field fw2">
            Sample
        </div>
        <div class="field fw5">
            Ingredient
        </div>
        <div class="field fw2">
            Quantity
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            1
        </div>
        <div class="field fw5">
            Pliag DNA
        </div>
        <div class="field fw2">
            5 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw5">
            10x buffer
        </div>
        <div class="field fw2">
            2 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw5">
            EcoRI
        </div>
        <div class="field fw2">
            1 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw5">
            Spel
        </div>
        <div class="field fw2">
            1 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw5">
            \(\mathrm{H_2 O}\) (MQ)
        </div>
        <div class="field fw2">
            11 µL
        </div>
    </div>
    
    <div class="record">
        <div class="field fw2">
            2
        </div>
        <div class="field fw5">
            Y4 DNA
        </div>
        <div class="field fw2">
            5 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw5">
            10x buffer
        </div>
        <div class="field fw2">
            2 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw5">
            Xbal
        </div>
        <div class="field fw2">
            1 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw5">
            EcoRI
        </div>
        <div class="field fw2">
            1 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw5">
            \(\mathrm{H_2 O}\) (MQ)
        </div>
        <div class="field fw2">
            11 µL
        </div>
    </div>
    </div>
    <div class="caption">Samples.</div>
</div>
<div class="text">Samples were digested for 2 hours at 37 °C. Samples were then loaded on 1% agarose gel with DNA stain G 1:50000</div>
<div class="object data">
    <div class="wrapper">
    <div class="header">
        <div class="field fw5">
            Ingredient
        </div>
        <div class="field fw3">
            Quantity
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
            sample DNA
        </div>
        <div class="field fw3">
            20 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
            6x buffer
        </div>
        <div class="field fw3">
            4 µL
        </div>
    </div>
    </div>
    <div class="caption">Samples.</div>
</div>
<div class="text">10 µL of NED 2log ladder. Gel was run on 100 V for 30 minutes. It was then cut.</div>
<div class="object data">
    <div class="wrapper">
    <div class="header">
        <div class="field fw2">
            Sample
        </div>
        <div class="field fw3">
            Weight
        </div>
        <div class="field fw4">
            Binding Buffer
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            1
        </div>
        <div class="field fw3">
            0.300 g
        </div>
        <div class="field fw4">
            600 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            2
        </div>
        <div class="field fw3">
            0.118 g
        </div>
        <div class="field fw4">
            236 µL
        </div>
    </div>
    </div>
    <div class="caption">Gel cuts.</div>
</div>
<div class="text">The samples were purified using a Thermo Scientific GeneJET PCR Purification Kit. Concentrations were checked using Nanodrop.</div>
<div class="object data">
    <div class="wrapper">
    <div class="header">
        <div class="field fw3">
            Sample
        </div>
        <div class="field fw3">
            Concentration
        </div>
    </div>
    <div class="record">
        <div class="field fw3">
            Pliag dig
        </div>
        <div class="field fw3">
            4,6 ng/µL
        </div>
    </div>
    <div class="record">
        <div class="field fw3">
            Y4 dig
        </div>
        <div class="field fw3">
            4,7ng/µL
        </div>
    </div>
    </div>
    <div class="caption">Sample concentrations.</div>
</div>
<div class="text">Samples were ligated overnight.</div>
<div class="object data">
    <div class="wrapper">
    <div class="header">
        <div class="field fw5">
        Ingredient
        </div>
        <div class="field fw2">
        Quantity
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
            Pliag DNA
        </div>
        <div class="field fw2">
            5 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
            Y4 DNA
        </div>
        <div class="field fw2">
            10 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
            10x T4 buffer
        </div>
        <div class="field fw2">
            2 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
            T4 Ligase
        </div>
        <div class="field fw2">
            0.5 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw5">
            \(\mathrm{H_2 O}\)
        </div>
        <div class="field fw2">
            0.5 µL
        </div>
    </div>
    </div>
    <div class="caption">Ligation mix.</div>
</div>
</html>
}} 

{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 7 August 2015 - 00:00, 7 August 2015
	|executor=Harm Ruesink
	|content=<html>
<div class="text">A heat shock transformation was performed. 20 µL of (Y4 + Pliag) DNA was added to 10 µL of competent NEB cells. This was plated on LB chloramphenicol plates and incubated overnight at 37 °C.</div>
</html>
}}

{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 8 August 2015 - 00:00, 8 August 2015
	|executor=Harm Ruesink
	|content=<html>
<div class="subtitle">Transformation results</div>
<div class="text"> No colonies were visible. </div>
</html>
}}

{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 9 August 2015 - 00:00, 9 August 2015
	|executor=Harm Ruesink
	|content=<html>
<div class="subtitle">Transformation results</div>
<div class="text">White colonies were visible. A colony PCR was performed. PCR showed possible insert in sample 9 and 13. Cultures were grown overnight to check for the insert. The overnight cultures were grown in LB + chloramphenicol (3mL) at  37 °C, 200rpm. </div>
</html>
}}

{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 11 August 2015 - 00:00, 11 August 2015
	|executor=Harm Ruesink
	|content=<html>
<div class="text">Cultures 9 and 13 were miniprepped and loaded on 1% agarose gel with DNA stain G 1:50000.</div>
<div class="object data">
    <div class="wrapper">
    <div class="header">
        <div class="field fw4">
            Ingredient
        </div>
        <div class="field fw2">
            Quantity
        </div>
    </div>
    <div class="record">
        <div class="field fw4">
            Sample DNA
        </div>
        <div class="field fw2">
            1 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw4">
            \(\mathrm{H_2 O}\)
        </div>
        <div class="field fw2">
            4 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw4">
            6x buffer
        </div>
        <div class="field fw2">
            1 µL
        </div>
    </div>
    </div>
    <div class="caption">Mix.</div>
</div>
<div class="text">10µL of Quick-Load® Purple 2-Log DNA Ladder (0.1 - 10.0 kb) was used. The gel was run at 100 V for 30 minutes. Nothing was visible on the gel. DNA concentration should be checked.</div>
</html>
}}

{{Team:Groningen/TEMPLATES/LOG
	|title=00:00, 13 August 2015 - 00:00, 13 August 2015
	|executor=Harm Ruesink
	|content=<html>
<div class="text">Concentrations were checked using nanodrop.</div>
<div class="object data">
    <div class="wrapper">
    <div class="header">
        <div class="field fw2">
            Sample
        </div>
        <div class="field fw5">
            Ingredient
        </div>
        <div class="field fw2">
            Quantity
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            9
        </div>
        <div class="field fw5">
            Sample DNA
        </div>
        <div class="field fw2">
            4 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw5">
            6x Buffer
        </div>
        <div class="field fw2">
            1 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw5">
            \(\mathrm{H_2 O}\)
        </div>
        <div class="field fw2">
            1 µL
        </div>
    </div>
    
    <div class="record">
        <div class="field fw2">
            13
        </div>
        <div class="field fw5">
            Sample DNA
        </div>
        <div class="field fw2">
            5 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw5">
            6x Buffer
        </div>
        <div class="field fw2">
            1 µL
        </div>
    </div>
    
    <div class="record">
        <div class="field fw2">
            Y4
        </div>
        <div class="field fw5">
            Sample DNA
        </div>
        <div class="field fw2">
            1 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw5">
            6x Buffer
        </div>
        <div class="field fw2">
            1 µL
        </div>
    </div>
    <div class="record">
        <div class="field fw2">
            
        </div>
        <div class="field fw5">
            \(\mathrm{H_2 O}\)
        </div>
        <div class="field fw2">
            4 µL
        </div>
    </div>
    </div>
    <div class="caption">Gel Samples.</div>
</div>
<div class="text">Gel was run at 100 V for 30 minutes. The gel showed no bands. </div>
</html>
}}