AlanFung:LabNotes/Reprogramming/2010-11-10

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Revision as of 22:30, 11 November 2010 by >Alan6017518 (→‎one step qRT-PCR)
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one step qRT-PCR

  • The purpose of this experiment is to perform quantitative RT-PCR for the detection of the efficiency of SPEN gene knockdown.


  • Superscript III RT/Platinum Taq Mix 1ul
  • 2X SYBR Green Reaction Mix 25ul
  • Forward Primer, 10uM 1ul
  • Reverse Primer, 10uM 1ul
  • Template (1pg to 1ug total RNA) ul
  • DEPC-treated water to 50ul
14 1.44 1.21
15 1.32 1.43
93 1.33 1.73
94 1.02 1.76
95 1.43 1.95
96 1.13 2.43
14 20.56 20.79
15 20.68 20.57
93 20.67 20.27
94 20.98 20.24
95 20.57 20.05
96 20.87 19.57
  • Running at Program “one step qrt_pcr"
  • 50C 3min -> hold -> 95C 5min -> hold (95C 15sec -> 60C 30sec) x 40 cycle -> 40C 1min