We have tried genotyping this line using Jackson's protocol and a protocol found online, but the gels are always blank (primers and ladders visible). No amplification of even the wildtype band (not even in a wild type control sample), even though primers map to exon 3 of the gene.
DNA prep is OK. Same samples genotype fine for the LyzM cre we bred in and a floxed gene we bred in (all on different chromosomes).
Have tried different Taqs and mastermixes.
Does anyone else use this line? Any trouble genotyping it? Any tricks to try?
hi
one thing to do is first test the primers for an in silico PCR (try on the UCSC or NCBI tools). if the in silico amplification is right, at the good size and position on genome, the PCR protocols are to be checked, but also quality of primers (false production?).
fred
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