Hi
I am struggling to design my primers for TAF15-201.I am supposed to do it for 2 consecutives exons, but I don’t know what I am supposed to do with introns Do I need the full intronic sequence? how do I link both exonic sequences?
Then once in my primer design tool, and knowing that I need to amplify at least 30bp upstream and downstream of the exons encoding TAF15-201 What are the ranges for my forward and reverse primer what is my pcr size ?
where do my primers sit in my sequence ?
Hi Barry
No You do not need the full intronic sequence unless you’re amplifying genomic DNA that includes intronic regions. You can design primers to amplify both exons and flanking regions (at least 30 bp upstream and downstream of each exon). Use the primer designing tool to input your target regions and find appropriate primer sequences. The PCR product size will be roughly the sum of the exonic sequences plus the 30 bp flanking each exon, e.g., for two 200 bp exons, the product might be ~460-500 bp.