Difference between revisions of "TSC-floxed Genotyping"
From Bridges Lab Protocols
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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. | 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. | ||
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| + | [[ Category:Genotyping ]] | ||
| + | [[ Category:Mouse Work]] | ||
| + | [[ Category:PCR]] | ||
| + | [[ Category:DNA]] | ||
| + | [[ Category:Molecular Biology]] | ||
| + | [[ Category:mTORC1 ]] | ||
Latest revision as of 19:11, 30 December 2012
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.
