Daniel:Notebook/ComboLock/2016-11-16: Difference between revisions

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| width="160" height="34" | Sample
| width="160" height="34" | Sample
| width="65" | ng/uL
| width="85" | ng/uL
| width="65" | ug in 30 uL
| width="85" | ug in 30 uL


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Revision as of 18:15, 16 November 2016

BSA Beads Test (Started Yesterday)

Back to Calendar

Protocol

  1. Rolling Circle Amplification
    1. Prepare 10.2X Master mix according to table below
    2. Add sample to new reaction tubes according to following table
    3. Reagent uL Added Master Mix (10.2X)
      Template 5 NA
      RCA Primer (100 uM) 2.5 25.5
      dNTP (1 mM) 0.8 8.16
      10X Buffer 2 20.4
      Phi29 1 10.2
      nfH2O 8.7 88.74
      Total 20 153
    4. Incubate at 37C for 3 hours
    5. Incubate at 65C for 10 minutes;Hold at 10C until next step
  2. qPCR

Library Prep (Started Yesterday; Exp. Start 11/7/16)

  1. Ethanol Precipitation
    1. Centrifuge for 25 minutes at 12000 rpm at 4C
    2. Remove supernatant and add 750 uL 70% EtOH
    3. Centrifuge for 12 minutes at 12000 rpm at 4C
    4. Remove supernatant and spin in vacuum centrifuge for 10 minutes
    5. Add 30 (20 for NTC) uL nfH2O to resuspend; pool samples A and B by reusing the water
Sample ng/uL ug in 30 uL
Cells RCA Product (2AB,2BB)