Matt:LabNotes/2013-5-10: Difference between revisions

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>Mzcai
(Created page with "==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...")
 
>Mzcai
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===Probe Expansion PCR===
===Probe Expansion PCR===
Expansion PCR in 2 tubes of 200ul total (1 tube for 0bp gap probes with AP1V6 primers; 1 tube for 20bp gap probes with AP1V4 primers) <br>
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) <br>
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94C 2min -> (94C 30sec -> 54C 45sec -> 72C 45sec) x 16 -> 72C 3min -> 15C hold
*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

Revision as of 00:18, 11 May 2013

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