Daniel:Notebook/ComboLock/2016-10-13

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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

Padlock0301 Dilution Test

Stock Dilutions

Create the following dilutions of PCC Template by taking 10 uL stock and diluting with 90 uL nfH2O (note:zero is normal 10 uM stock). #6 is the highest concentration, made on Friday 10-7.

Tube No Dilution Molecules Available Amount
6 1.00E+06 1.00E+06 10 amol
7 1.00E+07 1.00E+05 1 amol
8 1.00E+08 1.00E+04 100 zmol
9 1.00E+09 1000 10 zmol
10 1.00E+10 100 1 zmol
11 1.00E+11 10 100 ymol
12 1.00E+12 1 10 ymol

Buffers

Protocol