I would like to investigate association between this allele and a disease, but sequence of this allele is very similar with other alleles in hla_drb1 alleles group, I don't know how can I design a specific primer for this allele to not match to other alleles.
Using Primer 3 of NCBI, check the 'specificity check' under 'Primer Pair Specificity Checking Parameters'. But at first, you must have a specific sequence of the allele in fasta format.
The link of Primer 3 tool of NCBI:
https://www.ncbi.nlm.nih.gov/tools/primer-blast/index.cgi?LINK_LOC=BlastHome
Hi,
Adding dmso in increasing amounts until only the one primer can anneal or increasing the annealing temperature in a gradient (to see what temperature each forward primer anneals at without the other primers annealing) might be a solution.
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