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

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       <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols/yeast-dna-extraction-dneasy-blood-and-tissue-kit">
 
       <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols/yeast-dna-extraction-dneasy-blood-and-tissue-kit">
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         <p>Yeast DNA extraction using DNeasy® Blood & Tissue Kit [QIAGEN]</p>
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         <p>Yeast DNA extraction </p>
 
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       <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols/titration-acid-phytic">
 
       <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols/titration-acid-phytic">
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         <p>Titration of phytic acid using Megazyme© Kit</p>
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         <p>Titration of phytic acid</p>
 
         <img src="https://static.igem.org/mediawiki/2015/1/11/ParisBettencourt_protocolsGel.JPG">
 
         <img src="https://static.igem.org/mediawiki/2015/1/11/ParisBettencourt_protocolsGel.JPG">
 
       </a>
 
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Revision as of 12:04, 10 September 2015

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

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