Good qPCR primers for Nav1.8 and TRPV1

Experimental Techniques
1

Can you suggest confirmed qPCR primers and PCR program for Nav1.8 and TRPV1? Would be greatly appreciated. Thanks in advance, Vanja

2

Hi @vanja.

I can confirm that the sodium channel primers in this paper amplify the target efficiently:


I’m assuming your asking for Mouse.

@tberta Do you have a mouse TRPV1 primer?

3

PrimerBank from Harvard (https://pga.mgh.harvard.edu/primerbank/) is a free website and reliable resource for mouse and human primer sets. Most sets are validated and if they’re not we always select the ones close to the 5’ end of the transcript of interest (less sensitive to the RNAse degradation). For instance, we have successfully used the following TRPV1 primers from the PrimerBank: fw - CCGGCTTTTTGGGAAGGGT and rev - GAGACAGGTAGGTCCATCCAC.

The efficiency, specificity and sensitivity of the primer sets, as well as the qPCR program depend on the SYBR green mix used for the qPCR. We have good results with the PowerUp SYBR Green Master Mix from ThermoFisher.

4

Thanks Temo. I’ve had mixed results with Primer bank. But I’m glad to hear you used that primer and it worked for you.

5

With the exception of the primer sets for cytokines, we have a great success rate with PrimerBank. I would like to add that the the reverse transcription kit is also important to obtain good results. We like the QuantiTect Reverse Transcription Kit from Qiagen or the SuperScript™ VILO™ cDNA Synthesis Kit from ThermoFisher. They are pricey, but we generally reverse transcription in total volume of 10 ul, instead of the suggested total volume of 20 ul.

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