Matt:LabNotes/2016-7-26
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TB12k_Apr2016 V4 Probe Preparation
Production PCR
- Make 10ml V4 master mix for 96 wells
Components | Volume (1X) | Volume (100X) |
First round amplicon TB12k_Apr2016_V4 (10nM) | 0.2 | 20 |
2X KAPA SYBG MM | 50 | 5000 |
AP1V4U (100uM) | 0.4 | 40 |
AP2V4 (100uM) | 0.4 | 40 |
H2O | 49 | 4900 |
Total | 100 | 10,000 |
Program 98C 1min -> (98C 30sec -> 55C 45sec -> 72C 45sec) x 14 -> 72C 2min -> 4C hold
- Did in regular thermocyclers
EtOH Precipitation
- 8 15-ml tubes (with 12 wells (1.5 strips) of PCR product each) for
- 1,200ul PCR product
- 3000ul 100% EtOH
- 4ul GlycoBlue
- 120ul 3M NaOAc pH 5.2-5.5