Matt:LabNotes/2017-5-20

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Revision as of 19:42, 20 May 2017 by >Mzcai (Created page with "==Agi15k_Feb2017_V4 Probe Production== *Production of Padlock Probes (V4) Matt:LabNotes/2017-2-8 ===Notes from last time=== *Do 12 or 13 cycle...")
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Agi15k_Feb2017_V4 Probe Production

Notes from last time

  • Do 12 or 13 cycles Production PCR
  • Experiment with 1hr vs 2hr Lambda digestion
  • Think of way to maximize USER and DpnII digestion

Production PCR

Components Volume (1X) Volume (100X)
First round amplicon Agi15k_Feb2017_V4 (25nM) 0.1 10
2X KAPA SYBG MM 50 5000
AP1V4U (100uM) 0.4 40
AP2V4 (100uM) 0.4 40
H2O 49 4910
Total 100 10000

Program 98C 1min -> (98C 10sec -> 55C 30sec -> 72C 45sec) x 13 -> 72C 2min -> 15C hold