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