Noi/NOTES/2012-5-25

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Revision as of 22:53, 25 May 2012 by >Noi
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== PCR Amplification with barcoded primers (AmpF6.4Sol and AmpR6.3 IndX No.1-)

  • I tested if the capture works well by performing qPCR of few samples in small volume 25ul, sample E1, F1, G1 and NTC
Captured template 2.5 0
10uM AmpF6.4Sol 0.5 2.1
10uM AmpR6.3Sol 0.5 2.1
50x SYBG 0.2 0.84
2x Phusion MM 12.5 52.5
H2O 8.8 36.96
  • Aliquot 22.5ul to each tube and add 2.5ul of circularized DNA templates

Program

98C 30s -> (98C 10s -> 58C 20s -> 72C 20s)x8 -> (98C 10s -> 72C 20s)x15 -> 72C 3min
  • Form the qPCR of tested samples, I could observe amplification in every samples except NTC, so I can continue amplify all samples and barcode with index primers.