AlanFung:LabNotes/Capturing/2014-1-2

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Helping Dheeraj to test Klentaq capture reactions

  • Dheeraj reached out to me saying he wants to start a qPCR reaction on the Klentaq capture reaction, he was working with Noi to test the protocol, but Noi is sick and I will be helping to perform the qPCR
  • They have 12 samples and we will be amplifying the captured samples with qPCR using the same protocol as it in Stoffol capture
  • 1-6 done in condition 16 hours incubation 4 hours polymerization
  • the second set we did the same 20 hours incubation and 20 hours polymerization

Protocol

PCR amplification to check if capture works

Components 1x rxn (ul) 12x rxn (ul) Final Conc.
Circularized DNA template 2.5 0  
KAPA SSYBR FAST MM 2X 12.5 150 1X
10uM AmpF6.4Sol 0.5 6 200nM
10uM AmpR6.3IndX 0.5 6 200nM
H2O 10 120  
Total 25