Protocol 36278: Probe Assay - Banp<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

>chr8:121978231+121978363 133bp TTTTGGTTTTCAGGGTTTGG GCAATCTCACCTTTCCTGAG

Mut= 115 bp

Wt= 133 bp

Fam=Mut

Hex=Wt

Sequence

Wt Sequence (deletion in lower case):

 AGTTCCTGTGTGCGCCATGTACATGGTAGCTTGTGACAGGACTGGCCCTTGGGAGAGCAAAGCCATCCTGTTGAGCTTACCGAGCACATGGCCTGCTGCTGACTTCCAGCTTGCTAGTTGGAAACTGCTCCTCTTGTGGGCCACTAGGTGGCACACTTGATATTTAGAAGGTCCCTGGAAGGGGTCATTGACAGATTTCAGTGAGGTATAGCTGTGTCCAGCCATCTTTTTTTGGTTTTCAGGGTTTGGATAGTAGTCTTATCTTTCAGCATTAGAAAAATTTTGTAGCTTTACTGTataaatatttggatgagcatagtgacaagcttattactccatggcctcaggaaaggtgagattgcaagggcttattagacacagagggctgaatccctgtgcacggcccagggctcctcactgaatatgttggctcggcccagggctcactcagctgaaatacactgggtggacttaggagccagctagggtcctaaggacttgctgctcacagtggaagacatcctctctgaagcttttctgttttcatttccagtctttcctgtactctattaaccagacgatatgtttgcggttggatagcattgaggccaagctgcaagctctcgaggccacttgcaaatctctggaagagaagctagacctggtcaccaataaacagcacagtcccatccaggtccccatggtggcaggttccccacttggcgccacccagacctgcaacaaagtgcgatggtaagtccagaagaccgaggctccgcaggccaccctgggaaagcgctccgtgccagcccaggcgccgctgttactgatctgccctcgccccggATATCTGCCCTGGTGCTTCCATGGGGCTGGTTTCACTTAGACCGAAACCAAACACAGGCTCTGCTGGAACGGCCTGAGTTGCTCCAACTTTGTTTCTTGGCTTAAGCAGCTGGACTCATCTTTCCTTGCACCTGGCCATGAACTTGGCCCATGTATGTTGCTTCTTTTCTCTTTCTATCTCTGGCTGACTATTGCCTATGTTTTCTGCGGGGGTAATATTTATAATTTTTACTTGATCATAGAAAACTTTCCGTTTAGTCTGAGACTTGCCTTGTGCCTAATTT

This mutation is a 549 bp deletion beginning at Chromosome 8 position 121,978,299 bp and ending after 121,978,847 bp (GRCm38/mm10).

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
48288 TTT TGG TTT TCA GGG TTT GG Common A
48289 GCA ATC TCA CCT TTC CTG AG Wild type Reverse A
48290 TTT CGG TCT AAG TGA AAC CAG Mutant Reverse A
48291 Fluorophore-1 TTG GAT GAG CAT AGT GAC AAG C Quencher-1 WT Probe
48292 Fluorophore-2 AGC TTT ACT GTA TAT CTG CCC TGG T Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
48288 0.40 uM
48289 0.40 uM
48290 0.40 uM
Wt Probe 0.15 uM
Mutant Probe 0.15 uM
DNA

Cycling

Step Temp °C Time Note
1 95.0 --
2 95.0 --
3 60.0 --
4 -- repeat steps 2-3 for 40 cycles
5 40.0 -- Forever
JAX uses a very high speed Taq (~1000 bp/sec), use cycling times recommended for your reagents.

Strains Using This Protocol

This is the only strain that uses this protocol.