What are different ways of genotyping a transgenic mouse? - what is best way to do genotyping--PCR, qRT-PCR or RT-PCR (Apr/17/2008 )

The core facility in our university has injected the DNA for our transgenic mouse. They will be shipping some 23 potential founders.

can anybody please let me know what needs to be done next,

As far as I understand, after waiting for 6 weeks (when mouse are grown up ), I can take a tail piece and isolate DNA and then do genotyping .

Which is the best way to genotype, (a) simple PCR ( Real-Time PCR © Reverse-Transcriptase PCR and why ??

Do I need to screen all 23 founders??

I am new to this field, any help, suggestion is highly appreciated.
Thanks,
jhil

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No need to wait until they grow up. I take pieces of tail from younger mice and genotype them using normal PCR, one for the wt allele, one for the transgenic allele, and I always use wt DNA as a reference. You should ask the people who gave you the mice for a genotyping protocol; they should provide you with primer sequences and PCR conditions (although in my experience often the conditions need a bit of tweaking).

Ginger

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Oops. Just noticed that these are your own line, so scrap that last bit about asking for a procotol. You should at least have a PCR protocol in place to detect your sequence of interest, and I'd use plasmid DNA as a positive control and wt DNA as a negative control.

Good luck!

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Thanks for your help.

Whats the advantage of doing RT-PCR for genotyping ?

Can I do RT-PCR with the GENOMIC DNA isolated from transgenic positive mouse?

Thanks again !

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