AlanFung:LabNotes/Reprogramming/2010-11-10
Jump to navigation
Jump to search
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