Revision as of 14:06, 10 September 2015

Ferment It Yourself

iGEM Paris-Bettencourt 2O15

Analytical digestion protocol

Prepare the following mix:
- 2µL 10X Digestion buffer
- 0.5µL Eco31I
- 0.5µL BbsI
- 2µL of DNA (200ng)
- 15µL water
incubate 1h at 37°C

Annealing Protocol

Phosphorylation of the oligos

5.6μL DNAse/RNAse free water
6.0μL oligo 1 (10µM)
6.0μL oligo 2 (10µM)
2.0μL 10X T4 DNA ligase buffer
0.4μL T4 PolyNucleotide Kinase

Total: 20μL

incubate 30min at 37°C
add 1μL of 1M NaCl
incube 5min at 95°C
let the mix cool down
use 2μL of the mix as a 10X solution

Chemical test for competent cells

20-30 sec at 8-10 krpm for the DNA tube (from the kit).
Thow cell competent on ice. Label 2 ml eppendorf µtube for each concentration and put it on ice.
Add 1µl of DNA into each µtube.
Add 50µl of competent cell into each tube. Flick gently to mix. Incubate in ice for 30 minutes. Pre-heat waterbath at 42°C.
Heat-shock cell by placing in waterbath 1 minute. CAUTION : The top need to be close to the water level, but not immerge in.
Keep back the tube to ice for 5 minutes.
Add 200µl of SOC media, incubate 37°C for 2h. Prepare agar plate of the media that you want, and label it (3 plate for each concentration).
Add 20µl of each tube n the appropriate plate. Incubate 37°C for OVN (12-16h).

Analytical digestion

Annealing

Chemical test for competent cell

Electroporation

Electrocompetent Cells Preparation

Digestion

Electrocompetent Cells Preparation for Lactobacillus plantarum

Heat Shock transformation

Titration protocols for vitamin A

Titration protocols for vitamin A with HPLC

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Yeast lysis with NaOH

Heat shock transformation for yeast

Ligation

Titration protocols for vitamin B2 with HPLC

Idli recipe

Miniprep protocol

PCR purification

PCR

Gel purification

Yeast DNA extraction

Titration of phytic acid

Gel Electrophoresis with SYBR safe

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        <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols#AnalyticalDigestion">
-         <p>Analytical digestion</p>
+         <div class="ptext"><p>Analytical digestion</p>
          <img src="https://static.igem.org/mediawiki/2015/e/e3/ParisBettencourt_protocolsWhiteBoardAnalyticalDigestion.JPG">
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      <div class="innerBox cyan">
        <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols#Annealing">
-         <p>Annealing</p>
+         <div class="ptext"><p>Annealing</p></div>
          <img src="">
        </a>
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        <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols#ChemicalTestCompetentCells">
-         <p>Chemical test for competent cell</p>
+         <div class="ptext"><p>Chemical test for competent cell</p>
          <img src="https://static.igem.org/mediawiki/2015/e/ec/ParisBettencourt_protocolsHeatBlock.JPG">
        </a>
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        <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols/Electroporation">
-         <p>Electroporation</p>
+         <div class="ptext"><p>Electroporation</p></div>
          <img src="https://static.igem.org/mediawiki/2015/2/24/ParisBettencourt_protocolsCentrifuge.JPG">
        </a>
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        <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols/Electrocompetent Cells Preparation">
-         <p>Electrocompetent Cells Preparation</p>
+         <div class="ptext"><p>Electrocompetent Cells Preparation</p></div>
          <img src="https://static.igem.org/mediawiki/2015/d/da/ParisBettencourt_protocolsErlen.JPG">
        </a>
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      <div class="innerBox cyan">
        <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols/Digestion">
-         <p>Digestion</p>
+         <div class="ptext"><p>Digestion</p></div>
          <img src="https://static.igem.org/mediawiki/2015/6/6a/ParisBettencourt_protocolsTubesOnIce.JPG">
        </a>
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      <div class="innerBox magenta">
-       <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols/Electrocompetent Cells Preparation for Lactobacillus Plantarum">
+       <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols/Electrocompetent Cells Preparation for L. plantarum">
-         <p>Electrocompetent Cells Preparation for <i>Lactobacillus plantarum</i></p>
+         <div class="ptext"><p>Electrocompetent Cells Preparation for <i>Lactobacillus plantarum</i></p></div>
          <img src="https://static.igem.org/mediawiki/2015/e/ec/ParisBettencourt_protocolsFrozenEnzymes.JPG">
        </a>
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      <div class="innerBox yellow">
        <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols/Heat Shock transformation">
-         <p>Heat Shock transformation</p>
+         <div class="ptext"><p>Heat Shock transformation</p></div>
          <img src="https://static.igem.org/mediawiki/2015/0/0d/ParisBettencourt_protocolsBanana.JPG">
        </a>
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      <div class="innerBox cyan">
        <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols/Titration protocols for vitamin A">
-         <p>Titration protocols for vitamin A</p>
+         <div class="ptext"><p>Titration protocols for vitamin A</p>
        </a>
      </div>
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      <div class="innerBox cyan">
        <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols/Titration protocols for vitamin A with HPLC">
-         <p>Titration protocols for vitamin A with HPLC</p>
+         <div class="ptext"><div class="ptext"><p>Titration protocols for vitamin A with HPLC</p></div>
        </a>
      </div>
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      <div class="innerBox magenta">
        <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols/Yeast lysis with NaOH">
-         <p>Yeast lysis with NaOH</p>
+         <div class="ptext"><p>Yeast lysis with NaOH</p></div>
          <img src="https://static.igem.org/mediawiki/2015/c/ce/ParisBettencourt_protocolsTubesIncubator.JPG">
        </a>
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      <div class="innerBox yellow">
        <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols/Heat shock transformation for yeast">
-         <p>Heat shock transformation for yeast</p>
+         <div class="ptext"><p>Heat shock transformation for yeast</p>
          <img src="https://static.igem.org/mediawiki/2015/5/5e/ParisBettencourt_protocolsPipetting.JPG">
        </a>
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      <div class="innerBox cyan">
        <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols/Ligation">
-         <p>Ligation</p>
+         <div class="ptext"><p>Ligation</p></div>
          <img src="https://static.igem.org/mediawiki/2015/3/34/ParisBettencourt_protocolsPCR.JPG">
        </a>
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      <div class="innerBox magenta">
        <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols/Titration protocols for vitamin B2 with HPLC">
-         <p>Titration protocols for vitamin B2 with HPLC</p>
+         <div class="ptext"><p>Titration protocols for vitamin B2 with HPLC</p></div>
          <img src="https://static.igem.org/mediawiki/2015/1/1c/ParisBettencourt_protocolsTubeUnderHood.JPG">
        </a>
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      <div class="innerBox yellow">
        <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols/Idli receipe">
-         <p>Idli recipe</p>
+         <div class="ptext"><p>Idli recipe</p></div>
          <img src="https://static.igem.org/mediawiki/2015/5/55/ParisBettencourt_protocolsIdli.JPG">
        </a>
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      <div class="innerBox cyan">
        <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols/Miniprep protocol using a QIAGEN kit">
-         <p>Miniprep protocol</p>
+         <div class="ptext"><p>Miniprep protocol</p></div>
          <img src="https://static.igem.org/mediawiki/2015/e/ec/ParisBettencourt_protocolsQiagen.JPG">
        </a>
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      <div class="innerBox cyan">
        <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols/PCR purification">
-         <p>PCR purification</p>
+         <div class="ptext"><p>PCR purification</p></div>
          <img src="https://static.igem.org/mediawiki/2015/5/5f/ParisBettencourt_protocolsPCRtubes.JPG">
        </a>
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      <div class="innerBox cyan">
        <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols/PCR">
-         <p>PCR</p>
+         <div class="ptext"><p>PCR</p></div>
          <img src="https://static.igem.org/mediawiki/2015/3/34/ParisBettencourt_protocolsPCR.JPG">
        </a>
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      <div class="innerBox cyan">
        <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols/gel-purification">
-         <p>Gel purification</p>
+         <div class="ptext"><p>Gel purification</p>
          <img src="https://static.igem.org/mediawiki/2015/1/11/ParisBettencourt_protocolsGel.JPG">
        </a>
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        <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols/yeast-dna-extraction-dneasy-blood-and-tissue-kit">
-         <p>Yeast DNA extraction </p>
+         <div class="ptext"><p>Yeast DNA extraction </p></div>
          <img src="">
        </a>
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        <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols/titration-acid-phytic">
-         <p>Titration of phytic acid</p>
+         <div class="ptext"><p>Titration of phytic acid</p></div>
          <img src="https://static.igem.org/mediawiki/2015/1/11/ParisBettencourt_protocolsGel.JPG">
        </a>
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      <div class="innerBox cyan">
        <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols/Gel_Electrophoresis">
-         <p>Gel Electrophoresis with SYBR safe</p>
+         <div class="ptext"><p>Gel Electrophoresis with SYBR safe</p></div>
          <img src="https://static.igem.org/mediawiki/2015/1/11/ParisBettencourt_protocolsGel.JPG">
        </a>

Difference between revisions of "Team:Paris Bettencourt/Protocols"