Matt:LabNotes/2015-1-13: Difference between revisions
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>Mzcai (Created page with "==CA12k_Nov2014_V7 Probe Production== *Production of Padlock Probes targeting constitutive exon cDNA (V7 primers) of [[Matt:LabNotes/2014-11-19#Assembly_of_oligo_pool_for_orde...") |
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'''Program''' 98C 1min -> (98C 10sec -> 55C 30sec -> 72C 45sec) x 15 -> 72C 2min -> 15C hold | '''Program''' 98C 1min -> (98C 10sec -> 55C 30sec -> 72C 45sec) x 15 -> 72C 2min -> 15C hold | ||
[[File:011214_CA12kNov14_ProductionPCR_V7.JPG | 800px]] | |||
[[File: | 800px]] | |||
===EtOH Precipitation=== | ===EtOH Precipitation=== | ||
*6 15-ml tubes (with 8 wells of PCR product each) | *6 15-ml tubes (with 8 wells of PCR product each) for V7 | ||
**800ul PCR product | **800ul PCR product | ||
**2000ul 100% EtOH | **2000ul 100% EtOH |
Revision as of 04:21, 13 January 2015
CA12k_Nov2014_V7 Probe Production
- Production of Padlock Probes targeting constitutive exon cDNA (V7 primers) of Matt:LabNotes/2014-11-19#Assembly_of_oligo_pool_for_ordering
Production PCR
- 10nM 1st round amplicons
- V7 master mix
Components | Volume (1X) | Volume (50X) |
First round amplicon CA12k_Nov14_V7 (10nM) | 0.2 | 10 |
2X KAPA SYBG MM | 50 | 2500 |
AP1V7U (100uM) | 0.4 | 20 |
AP2V7 (100uM) | 0.4 | 20 |
H2O | 49 | 2450 |
Total | 100 | 5000 |
Program 98C 1min -> (98C 10sec -> 55C 30sec -> 72C 45sec) x 15 -> 72C 2min -> 15C hold File:011214 CA12kNov14 ProductionPCR V7.JPG
EtOH Precipitation
- 6 15-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