Hi,
We purchased tm1b mice where homozygous mutant is lethal so we have Het and corresponding WT.
When i am trying to geneotype i see a higher band indicating presence of distal LoxP site in all the cases. Does that mean the WT also contains the targeted allele? If i want to identify mutant, just the presence of LacZ indicate my protein is mutated? or only absence of exon means protein is mutated.
Thanks
does your no template control also show this band?
If so you have a contamination problem
You have to look where exactly your primers are located, if you bought the mice from KOMP then presence of LacZ means your protein is mutated as the lacz will make a frame shift mutation lead to early stop codon as it happens with tm1a mice.
For tm1b mice you can get it by crossing tm1a with Cre mice so the floxed exons will be removed leading to a null mutation.
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