I want to do RACE and I don't have any previous exposure. I am planning on using RLM-RACE kit invitrogen. But I am confused in a few places:
1. Should I choose the 5' or 3'? What criteria should I base my decision on?
2. After use (5' or 3') how do you store the product?
3. For sequencing do I clone it? Or do I give the product directly? I read that for the NGS-based approach there is no need for cloning.
Hello, see you can begin with 5'or 3' RACE no issues, but i suggest you should start with 3'RACE as any 3'region of a gene is more conserved so comparatively easier than 5'RACE.
You have to attempt both 5' and 3' RACE to get full-length of your gene. store all your RACE PCR products at -20C. Also, you may clone the putative fragment and perform sequencing reactions for clone confirmation.
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