AlanFung:LabNotes/Reprogramming/2010-11-10: Difference between revisions

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==one step qRT-PCR==
==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.
*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
*Running at Program “one step qrt_pcr"
*Running at Program “one step qrt_pcr"
*50C 3min -> hold -> 95C 5min -> hold (95C 15sec -> 60C 30sec) x 40 cycle -> 40C 1min
*50C 3min -> hold -> 95C 5min -> hold (95C 15sec -> 60C 30sec) x 40 cycle -> 40C 1min

Revision as of 23:20, 9 November 2010

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


  • Running at Program “one step qrt_pcr"
  • 50C 3min -> hold -> 95C 5min -> hold (95C 15sec -> 60C 30sec) x 40 cycle -> 40C 1min