Matt:LabNotes/2015-1-5

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Revision as of 20:39, 5 January 2015 by >Mzcai (Created page with "==CA12k_Nov2014 Probe Production== *Production of Padlock Probes targeting cDNA (V4 and V7 primers) of Matt:LabNotes/2014-11-19#Assembly_of_oligo_pool_for_ordering ===Exp...")
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CA12k_Nov2014 Probe Production

Expansion PCR

  • Matt:LabNotes/2014-12-18 conclusion: 21 cycles of PCR starting with 100nM seed oligos
  • Make 2 tubes:
    • V4 primers - CA12k_Nov2014_Contig
    • V7 primers - CA12k_Nov2014_Const
' Contig of Exons (V4) Constitutive Exons (V7)
CA12k_Nov2014 Seed Oligos (900nM) 15 15
2X KAPA SYBG MM 67.5 67.5
100uM AP1V4U 0.6 0
100uM AP2V4 0.6 0
100uM AP1V7U 0 0.6
100uM AP2V7 0 0.6
H2O 51.3 51.3
Total 135 135

Program 95C 30sec -> (95C 5sec -> 55C 30sec-> 72C 45sec) x 21 -> 72C 2min -> 15C hold

  • Purified with Qiagen columns

Production PCR