TSC-floxed Genotyping

From Bridges Lab Protocols
Revision as of 19:10, 30 December 2012 by Davebrid (talk | contribs) (Added primers and reference for TSC1 primers)
(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)
Jump to navigation Jump to search
The printable version is no longer supported and may have rendering errors. Please update your browser bookmarks and please use the default browser print function instead.

Primers

  • Cre (F) GCATT ACCGG GCAAC GAGTG ATGAG
  • Cre (R) GAGTG AACGA ACCTG GTCGA AATCA GTGCG
  • TSC1 (F) AGG AGG CCT CTT CTG CTA CC
  • TSC1 (R) CAG CTC CGA CCA TGA AGT G

Reference

From Kwiatkowski et al., Hum. Mol. Genet. 11(5):525-34. 2002. A mouse model of TSC1 reveals sex-dependent lethality from liver hemangiomas, and up-regulation of p70S6 kinase activity in Tsc1 null cells. http://www.ncbi.nlm.nih.gov/pubmed/11875047:

PCR genotyping was performed by simultaneous amplification of both wild-type Tsc1 and the deleted allele using the following three primers in a 35 cycle PCR reaction using Perkin Elmer AmpliTaq Gold: F4536, 5′-AGGAGGCCTCTTCTGCTACC-3′; R4830, 5′-CAGCTCCGACCATGAAGTG-3′; and R6548, 5′-TGGGTCCTGACCTATCTCCTA-3′ (Fig. 1D). Products were 295 bp (wild-type) and 368 bp (mutant), and were analyzed on agarose gels.

Retrieved from "http://bridgeslab.sph.umich.edu/protocols/index.php?title=TSC-floxed_Genotyping&oldid=736"