Team:TU Darmstadt/labbook

From 2013.igem.org

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                         </li></ul></ul>
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                         <ul><li>preparative 1% agarose gel
                         <ul><li>preparative 1% agarose gel
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                                 <ul><li>gel displays bands of expected size ' assembly was successful</li>
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                                 <ul><li>gel displays bands of expected size, therefore assembly was successful</li>
                         </ul></li></ul>
                         </ul></li></ul>
                 </td>
                 </td>

Revision as of 11:24, 3 October 2013

Labbook | Protocols | Materials |

13.08 gBLOCK assembly of CMK and TLO
  • PCR mixture
    • 1 µL of each gBLOCK
      • CMK: B_01 - B_04
      • TLO: A_01 - A_06
    • 10 µL 5x Q5 Reaction Buffer
    • 2 µL dNTPs
    • 1 µL Q5 Hot Start Polymerase
    • 10 µL 5x Q5 High GC Enhancer
    • 1 µL primer suffix-R (10 mM)
    • 1 µL primer prefix_R (10 mM)
  • PCR program (40 cycles)
    • initial denaturation 94°C, 100s
    • denaturation 94°C, 55s
    • annealing 64°C, 55s
    • elongation 72°C, 120s
    • final elongation 72°C, 300s
  • preparative 1% agarose gel
    • gel displays bands of expected size, therefore assembly was successful

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