I want to quantify gene expression from animal tissue using RT-PCR.I have some primers. The primers are used with a sybr green probe.I also have B-actin but it is used with a taqman probe.Can I use this bactin (endogenous control) and other primers same plate? Will they work?
- No
- Do not mix Taqman probes with cold primers binding Sybr green
- Choose your assay: Either purchase further Taqman probes or make ordinary primers to b actin and then perform sybr green qPCR
I agree with Laurence Stuart Dawkins-Hall that Taqman probes aren't designed to work with cold primers binding SYBR Green. The cheaper route to design your assay would be making ordinary primers to beta actin and then performing the SYBR Green qPCR. However, a more efficient reaction (but more expensive) assay would be designing Taqman probes for this assay. Just a side note, ensure that the endogenous control you are using has been proven to be a highly stable endogenous control within your animal tissue type because reference genes have been proven to not be stable nor ubiquitous between tissue types.
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