Rui:LabNotes/SingleCell/2011-5-21

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Revision as of 17:14, 23 May 2011 by >RuiLiu (Created page with '==Repeat 2nd strands and PCR with the remaining samples== ===QPCR curve=== ===Beads purification=== * Combined into 4 samples, 8 PCR tubes per samples * Sperm RNA, ES RNA, Sper…')
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Repeat 2nd strands and PCR with the remaining samples

QPCR curve

Beads purification

  • Combined into 4 samples, 8 PCR tubes per samples
  • Sperm RNA, ES RNA, Sperm lysis, ES lysis
  • 0.7X beads purification
  • Elute with 50ul water
Sample ID ng/ul 260/280 260/230
sperm RNA 43.64 1.9 1.68
ES RNA 295.67 1.93 2.18
ES lysis 220.08 1.92 1.97
sperm lysis 23.5 1.68 0.95

Cloning

  • Ligation: ~25-30ng purified PCR + 2.5ng vector in 10ul total ligation, 14C, o/n
  • Transformation, using Kan30 plate