Noi/NOTES/2011-10-6
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Plan of probe (from LC Bioscience) production
- Follow the protocol: [1]
- After receiving the probes, resuspend with H2O to the concemtration 20 nM
- perform expansion PCR to amplify the probe as the template
Components | Volume (ul) | Final conc. |
Resuspended probes (100nM) | 5.00 | 2.5nM |
eMIP_CA1_F (100uM) | 0.80 | 400nM |
eMIP_CA1_F (100uM) | 0.80 | 400nM |
2x Phusion master mix | 100.00 | 1x |
H2O | 93.40 | |
Total | 200.00 |
Program
98C 30s -> (98C 10s -> 55C 20s -> 72C 20s)x16 -> 72C 5 min -> 15C hold
- Purified with Qaiquick column (2 columns) and elute with EB buffer volume 50ul each -> total volume 100ul.
- Measure DNA conc. with Nanodrop and adjust conc. to 100nM