AlanFung:LabNotes/Capturing/2012-6-22
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- Perform one step qPCR using Indexing seq. primer
Content | Volume | MM |
Captured DNA | 10 | Do not add to MM |
2X Kapa SYBG qPCR master mix | 50 | 110 |
10uM AmpF6.3Sol | 4 | 8.8 |
10uM AmpR6.3Ind | 4 | Do not add to MM |
H2O | 32 | 70.4 |
98C 30S -> (98C 10S -> 58C 20S -> 72C 20S) x 6 -> (98C 10S -> 72C 20S) x40
Monitoring the reactions and terminate the program right before the amplification curves reach the plateau. Ind1-H7gDNA Ind2-H14gDNA
File:ZhangLab 2 2012-06-23 18hr 06min 1-6.jpg
- Calculate amplicon size
AmpF6.3Sol AATGATACGGCGACCACCGACACTCTCAGATGTTATCGAGGTCCGAC AmpR6.3Ind1 CAAGCAGAAGACGGCATACGAGATCGTGATGCTAGGAACGATGAGCCTCCAAC