Matt:LabNotes/2015-2-2: Difference between revisions
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>Mzcai (Created page with "==CA12k_Nov2014_V4 Probe Production== *Production of Padlock Probes (V4) Matt:LabNotes/2014-11-19#Assembly_of_oligo_pool_for_ordering *Do double of what I normally do (96 ...") |
>Mzcai m (→Production PCR) |
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| 2X KAPA SYBG MM||50||5000 | | 2X KAPA SYBG MM||50||5000 | ||
|- | |- | ||
| | | AP1V4U (100uM)||0.4||40 | ||
|- | |- | ||
| | | AP2V4 (100uM)||0.4||40 | ||
|- | |- | ||
| H2O||49||4900 | | H2O||49||4900 |
Revision as of 22:23, 2 February 2015
CA12k_Nov2014_V4 Probe Production
- Production of Padlock Probes (V4) Matt:LabNotes/2014-11-19#Assembly_of_oligo_pool_for_ordering
- Do double of what I normally do (96 100ul PCR reactions instead of 48)
Production PCR
- 10nM 1st round amplicons
- V4 master mix made in 2 5ml tubes
Components | Volume (1X) | Volume (100X) |
First round amplicon CA12k_Nov14_V7 (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 | 10000 |
Program 98C 1min -> (98C 10sec -> 55C 30sec -> 72C 45sec) x 15 -> 72C 2min -> 15C hold [[File: | 800px]]