Table 7 Pipetting scheme for sgRNA amplification PCR.
From: An improved method for high-throughput quantification of autophagy in mammalian cells
Component | Amount per reaction (µl) | Final concentration |
|---|---|---|
NEBNext High Fidelity PCR Master Mix, 2 ×  | 25 | 1x |
Pooled sgRNA library template |  | 10.4 ng/µl |
NGS-Lib-Fwd primer (unique), see Table 9 | 1.25 | 0.25 µM |
NGS-Lib-KO-Rev, see Table 9 | 1.25 | 0.25 µM |
Pyrogen/RNase/DNase free water |  | add to a total volume of 50 µl |
