Daniel:Notebook/ComboLock/2016-10-13: Difference between revisions

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(Created page with "=Padlock0301 Test (Started Friday 10-7)= Back to Calendar ==Library Prep== <ol start="2"> <li>Size Sel...")
 
>Djacobse
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<li>Remove supernatant and add 750 uL 70% EtOH</li>
<li>Remove supernatant and add 750 uL 70% EtOH</li>
<li>Centrifuge for 12 minutes at 12000 rpm at 4C</li>
<li>Centrifuge for 12 minutes at 12000 rpm at 4C</li>
<li>Remove supernatant and spin in vacuum centrifuge for 10 minutes<li>
<li>Remove supernatant and spin in vacuum centrifuge for 10 minutes</li>
<li>Add 30 (20 for NTC) uL nfH2O to resuspend; pool samples A-C by reusing the water</li>
<li>Add 30 (20 for NTC) uL nfH2O to resuspend; pool samples A-C by reusing the water</li>
</ol></ol>
</ol></ol>

Revision as of 17:24, 13 October 2016

Padlock0301 Test (Started Friday 10-7)

Back to Calendar

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