Protocol 45764: Probe Assay - Eif4a2<em1(IMPC)J>
Version 1.0

Notes

Taqman qPCR protocols are run on a real time PCR instrument. Use an appropriate instrument specific Fluorophore/Quencher combination.

 

The genotyping protocol(s) presented here have been optimized for reagents and conditions used by The Jackson Laboratory (JAX). To genotype animals, JAX recommends researchers validate the assay independently upon receipt of animals into their facility. Reaction cycling temperature and times may require additional optimization based on the specific genotyping reagents used.

Expected Results

chr16:23108407+23108548 142bp GTCCAAGTCCCAAGCTGTTG AGCCACGCAGAGACCCTTA

Mutant=  137 bp

Wild Type = 142 bp

Fam = Mut

Hex = Wt

Sequence

Wt Sequence (deletions in lower case):

TCGGCGCGCGGAGCACTCGCGAGAGCTTCGCTTCCCAAGCGGGAGGGCCTGGTCCTAGGGCACTGAGTAAAGGGGCGTCTATACTTAGGGGTCCAAGTCCCAAGCTGTTGGGGAGGGCGAATGGCGCTTCGCAGCATTGCGGCCGGTTTTGAGGCGTTTTCAGCAGAGCacaggggcgcggaggggaatgaggtacggggtggaagggccctttaagggtctctgcgtggcttggaagcagtatgtctaactgcagtgttgttgtcagcagagaacatggcggcccagagggaatggaccccgatggtgtcatcgaggtaaggagtgggtgtggacgttaaaggtttggaaaggtacatttttttttcatgtcttattcatagtttcattattaacttacagagcaactggaatgaaattgttgataactttgatgatatgaatttaaaggagtcccttcttcgaggcatctatgcatatggttttgagaagccttcagctattcagcaaagagctattatcccttgtattaaaggtaaaactctattgccgttaggataagtgctctgcacatacatggaccggaaaagggaaagtgacctttggggaagtactagcacctgattgtattccttaaagtgaaatgatggcaaatcatctttcgggactgacctgaaatgaagagaatactcttgctgatcacttatgtgggcgtaactttatgttccaactggaatacatggtgtatagtagaagtaacgatttgatgtgggatcggtaaatatgtatcctactttttttagggtatgatgtgattgctcaagctcagtcaggtactggcaagacagccacatttgctatttccatcctgcaacagttggagattgagttcaaggagacccaagcactagtattggcccccaccagagaactggctcaacaggtattgatagtgtagttcaaaaaactgcttgcgtgttgcataggtttcaggtttcacaactgtgaagaatttaaaattaaaacgtataaattgtcctagtcctgtgagagcccccattttaattgtccatgcatgcagggagttttgttgaaagctaagcaggaaatgggtattgtaggtatgactttagggattgattgcacctcaacagttaaacccatAGGTTTCTCAGGGTGGATTATATCTGTACCCATTTAAAATTTGGAATGCAGTTGTGCAACATGAAAACTGCAGTGACATGTTAAACATTTGACTGTCTCAGTAGTTTGTGATGCATCTGTTGCATGCTATGTTTTCAAAGCACTGCTATATTGGCTTTGAAGTAATACTTCCTGATGAAGCTGCAAGCTTTATTCG

This mutation is a 995 bp deletion beginning at Chromosome 16 position 23,108,486 bp and ending after 23,109,480 bp (GRCm38/mm10).

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
66408 GTC CAA GTC CCA AGC TGT TG Common
66409 AGC CAC GCA GAG ACC CTT A Wild type Reverse
66410 TGC ACA ACT GCA TTC CAA A Mutant Reverse
66411 Fluorophore-1 CAG CAG AGC AGG TTT CTC AGG G Quencher-1 MUT Probe
66441 Fluorophore-2 TTC CAC CCC GTA CCT CAT T Quencher-2 WT Probe

Strains Using This Protocol

This is the only strain that uses this protocol.