Noi/NOTES/2011-10-6

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Revision as of 23:09, 6 October 2011 by >Noi (Created page with '== Plan of probe (from LC Bioscience) production == * Follow the protocol: [http://arep.med.harvard.edu/pdf/Porreca07.pdf]<br> * After receiving the probes, resuspend with H2O to…')
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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