Hi, everyone,
Thank you for your attention,
A question have being confused me deeply: How to design the qRT-PCR primer (SYBR Green) to detect the expression of A-T rich gene?
What the extent of A-T rich of the interested gene, I' d like to tell you, it like the repeat sequences, eg LINE1.
Thank you again, and looking forward to your answer.
Your help are need!
Hello! There was a Q&A opened a while ago for something similar. You might want to check it out.
https://www.researchgate.net/post/How_to_design_primers_from_AT_rich_region
Also, it is important to mention if you have the target sequence of the gene.
You could, as well, verify if there are readily available primers for your gene of choice.
This might help as well to design your primers: https://bitesizebio.com/10041/designing-qpcr-primers/
https://www.researchgate.net/post/How_to_design_Real_time_PCR_primers
And this article:
Article qPCR primer design revisited
Best of luck!
You can design a TaqMan Probe inside the AT rich region of your gene of interest and regular primers outside of the AT rich zone for qPCR. TaqMan gives you flexibility to design longer primers also.
Thank you very much @Ankur Bothra,
I knew TaqMan would be a better choice for this situation, it is truly more specific, however, it is also more expensive.
That's why I am trying to find out whether I can use SYBR green.
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