Matt:LabNotes/2015-1-5
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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
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