Matt:LabNotes/2015-2-5

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Revision as of 00:21, 6 February 2015 by >Mzcai (Created page with "==CA12k_Nov2014_V7 Probe Production== *Production of Padlock Probes (V7) Matt:LabNotes/2014-11-19#Assembly_of_oligo_pool_for_ordering *Do double of what I normally do (96 ...")
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CA12k_Nov2014_V7 Probe Production

Production PCR

Components Volume (1X) Volume (100X)
First round amplicon CA12k_Nov14_V7 (10nM) 0.2 20
2X KAPA SYBG MM 50 5000
AP1V7U (100uM) 0.4 40
AP2V7 (100uM) 0.4 40
H2O 49 4900
Total 100 10000

Program 98C 1min -> (98C 10sec -> 55C 30sec -> 72C 45sec) x 15 -> 72C 2min -> 15C hold [[File: | 800px]]

EtOH Precipitation

  • 12 5-ml tubes (with 8 wells of PCR product each) for V7
    • 800ul PCR product
    • 2000ul 100% EtOH
    • 2.7ul GlycoBlue
    • 80ul 3M NaOAc pH 5.2-5.5
  • Vortexed and put in -80C for overnight