Daniel:Notebook/ComboLock/2016-10-13

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Revision as of 17:57, 13 October 2016 by >Djacobse (→‎Library Prep)
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Padlock0301 Test (Started Friday 10-7)

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Library Prep

  1. Size Select Gel
    1. Centrifuge tubes at 12000 rpm for 1.5 minutes to bring gel to bottom
    2. Extract supernatant and add to nanosep column
    3. Centrifuge at 12000 rpm for 1.5 minutes; collect supernatant
  2. Ethanol Precipitation
    1. Add 1250 uL 100% EtOH, 50 uL 3M NaOAc, and 1.5 uL glycoblue to each sample
    2. Incubate at -80C for a while
    3. Centrifuge for 25 minutes at 12000 rpm at 4C
    4. Remove supernatant and add 750 uL 70% EtOH
    5. Centrifuge for 12 minutes at 12000 rpm at 4C
    6. Remove supernatant and spin in vacuum centrifuge for 10 minutes
    7. Add 30 (20 for NTC) uL nfH2O to resuspend; pool samples A-C by reusing the water