Matt:LabNotes/2013-5-10

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Revision as of 00:18, 11 May 2013 by >Mzcai (→‎Probe Expansion PCR)
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Agilent Probe Prep

  • Dr. Zhang resuspended probes to 20nM
  • Dr. Zhang aliquoted Agilent probes into separate tube labelled "FISHYB V1 probes 5/07/13"

Probe Expansion PCR

Expansion PCR in 2 tubes of 180ul total (1 tube for 0bp gap probes with AP1V6 primers; 1 tube for 20bp gap probes with AP1V4 primers)

' 0 gap probes (ul) 20 gap probes (ul)
Agi26k (20nM) 10 10
2x Kapa SYBG qPCR MM 100 100
100uM pAP1V6U 0.8 0
100uM AP2V6 0.8 0
100uM pAP1V4U 0 0.8
100uM AP2V4 0 0.8
H2O 88.4 88.4
Total 200 200
  • NOTE: Mixed thoroughly and then removed 20ul so there would be room to close lid

94C 2min -> (94C 30sec -> 54C 45sec -> 72C 45sec) x 14 -> 72C 3min -> 15C hold

  • Stopped after 12 cycles