Difference between revisions of "Team:SPSingapore/Notebook-Week-4"

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<font style = "color:blue;font-weight:bold;font-size:11pt;">&#9728; Yi Han &#9728;</font>
 
<font style = "color:blue;font-weight:bold;font-size:11pt;">&#9728; Yi Han &#9728;</font>
 
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<br><br>
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<span class = "nbcontent">
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&#10097;&#10097;&#10097; Received 2 bacterial stab cultures, EsaR/I plasmid from addgene (CHL) and BBa_K299812 from iGEM HQ.
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Received 2 bacterial stab cultures, EsaR/I plasmid from addgene (CHL) and BBa_K299812 from iGEM HQ.
 
</span>
 
</span>
 
<br>
 
<br>
 
<span class = "nbcontent">
 
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&#10097;&#10097;&#10097; Streaked out on plates with amp.
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Streaked out on plates with amp.
 
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</ul>
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<font style = "color:blue;font-weight:bold;font-size:11pt;">&#9728; Adrian &#9728;</font>
 
<font style = "color:blue;font-weight:bold;font-size:11pt;">&#9728; Adrian &#9728;</font>
 
<br><br>
 
<br><br>
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&#10097;&#10097;&#10097; Transformed:<br>
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&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;+Kit plate 1 9N Ba_K763002 chl<br>
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&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;+Kit plate 4 13L BBa_E0040 amp
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Transformed:
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<br>
 
<br>
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&#10097;&#10097;&#10097; DNa was received in powder form in plates, and resuspended in 10ul ultrapure H2O respectively. Plates were stored in -20/
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<span>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;+Kit plate 1 9N Ba_K763002 chl</span><br>
 +
<span>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;+Kit plate 4 13L BBa_E0040 amp</span>
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<br>
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<span class = "nbcontent"> DNa was received in powder form in plates, and resuspended in 10ul ultrapure H2O respectively. Plates were stored in -20/<span>
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</div>
 
<br><br>
 
<br><br>
 +
 
<font style = "color:blue;font-weight:bold;font-size:11pt;">&#9728; Yi Han &#9728;</font>
 
<font style = "color:blue;font-weight:bold;font-size:11pt;">&#9728; Yi Han &#9728;</font>
 
<br><br>
 
<br><br>
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&#10097;&#10097;&#10097; Plates cracked in incubator as they dried from lack of humidity.
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<div class = "divnbcontent">
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<span class = "nbcontent"> Plates cracked in incubator as they dried from lack of humidity.</span>
 
<br>
 
<br>
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&#10097;&#10097;&#10097; Transfer to small incubator with beaker of water for humidity no single colonies for inv plasmid -> streak again.
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<span class = "nbcontent"> Transfer to small incubator with beaker of water for humidity no single colonies for inv plasmid -> streak again.</span>
 
<br>
 
<br>
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&#10097;&#10097;&#10097; Wrong antibiotic for EsaR/I plasmid &rarr; streak out again
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<span class = "nbcontent"> Wrong antibiotic for EsaR/I plasmid &rarr; streak out again </span>
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<br><br>
 
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<br>
 
<br>
 
&#10097;&#10097;&#10097; Transformation of 13L repeated with 1ul of DNA.
 
&#10097;&#10097;&#10097; Transformation of 13L repeated with 1ul of DNA.
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<font style = "color:blue;font-weight:bold;font-size:11pt;">&#9728; Xin Yi &#9728;</font>
 
<font style = "color:blue;font-weight:bold;font-size:11pt;">&#9728; Xin Yi &#9728;</font>
 
<br><br>
 
<br><br>
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&#10097;&#10097;&#10097; No colonies grew for 13L on all plates &rarr; adjust incubator, make new media
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<div class = "divnbcontent">
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<span class = "nbcontent"> No colonies grew for 13L on all plates &rarr; adjust incubator, make new media</span>
 
<br><br>
 
<br><br>
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&#10097;&#10097;&#10097; Miniprep of inoculated bacteria for invasin plasmid, incubate sample in 37degC for 1h 20min:
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<span class = "nbcontent"> Miniprep of inoculated bacteria for invasin plasmid, incubate sample in 37degC for 1h 20min:</span>
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<br><br>
 
<br><br>
 
&#10097;&#10097;&#10097; Restriction digest for EsaR with XbaI/BamHI:
 
&#10097;&#10097;&#10097; Restriction digest for EsaR with XbaI/BamHI:
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<br><br>
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<br>
 +
<div style = "margin-left: 50px">
 
<span><b><u>RE reaction</u></b></span><br>
 
<span><b><u>RE reaction</u></b></span><br>
 
<span>5ul plasmid</span><br><span>5ul buffer</span><br><span>1ul XbaI</span><br><span>1ul BamHI</span><br><span>Add H2O to 50ul</span>
 
<span>5ul plasmid</span><br><span>5ul buffer</span><br><span>1ul XbaI</span><br><span>1ul BamHI</span><br><span>Add H2O to 50ul</span>
 +
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<br><br>
 
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Revision as of 18:18, 13 September 2015


Research Notebook

Week 1 (24/5 - 30/5)

May 25

☀ Yi Han ☀

Received 2 bacterial stab cultures, EsaR/I plasmid from addgene (CHL) and BBa_K299812 from iGEM HQ.
Streaked out on plates with amp.


May 26

☀ Adrian ☀

Transformed:
      +Kit plate 1 9N Ba_K763002 chl
      +Kit plate 4 13L BBa_E0040 amp
DNa was received in powder form in plates, and resuspended in 10ul ultrapure H2O respectively. Plates were stored in -20/


☀ Yi Han ☀

Plates cracked in incubator as they dried from lack of humidity.
Transfer to small incubator with beaker of water for humidity no single colonies for inv plasmid -> streak again.
Wrong antibiotic for EsaR/I plasmid → streak out again


May 28

☀ Yi Han ☀

❱❱❱ Inoculate single colony of invasin plasmid carrying bacteria in 3mL LB+amp
❱❱❱ Transformation of 13L repeated with 1ul of DNA.

May 29

☀ Xin Yi ☀

No colonies grew for 13L on all plates → adjust incubator, make new media

Miniprep of inoculated bacteria for invasin plasmid, incubate sample in 37degC for 1h 20min:

RE of invasin plasmid RE control
Rsal 1ul Rsal 0 ul
EcoRI 1 ul EcoRI 0 ul
INv plasmid 7.5ul Inv plasmid 7.5 ul
H2O 35.5 ul H2O 37.5 ul
NEB buffer 4.5 ul Buffer 4.5 ul

❱❱❱ Repeat transformation of 13L with 1ul
May 30

☀ Xin Yi & Yi Han ☀

❱❱❱ Miniprep of EsaR/I plasmid for 4 colonies

❱❱❱ Restriction digest for EsaR with XbaI/BamHI:
RE reaction
5ul plasmid
5ul buffer
1ul XbaI
1ul BamHI
Add H2O to 50ul