Matt:LabNotes/2017-3-30
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Agilent 15k Feb2017 Oligos
- Probe design
- Oligonucleotide Library received (Part Number G7221A)
- Dan Konet says the pools yield ~10pmols
- I resuspended to 200ul with nf-H2O (50nM concentration if actually 10pmols)
Probe Expansion PCR
- Expansion PCR in 4 tubes of 100ul
- V4 primers for Human Brain and V7 primers for Mouse Brain
' | 0 gap probes (ul) | 20 gap probes (ul) |
Agi15k (50nM) | 10 | 10 |
2x Kapa SYBG qPCR MM | 100 | 100 |
100uM pAP1V7U | 0.8 | 0 |
100uM AP2V7 | 0.8 | 0 |
100uM pAP1V4U | 0 | 0.8 |
100uM AP2V4 | 0 | 0.8 |
H2O | 88.4 | 88.4 |
Total | 200 | 200 |
- NOTE: Mixed thoroughly and then removed 20ul so there would be room to close lid
94C 2min -> (94C 30sec -> 54C 45sec -> 72C 45sec) x 14 -> 72C 3min -> 15C hold