Protocol 37081: Probe Assay - Snx12<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

>chrX:101214535-101214634 100bp GAAATGGAGGGAAGGAAGGA AATAGCAGCCCAGTTGGAGA

Mut = 102 bp

Wt = 100 bp

Fam = Mut

Hex = Wt

 

Sequence

Wt Sequence (deletions in lower case):

ACAGGATTATTAAAGAAAGTGAAGAGTACTTGAAGTTTATTTCCTTCATTGAGATAGTAACTCTGTCCTTCCACAGAAATCAAGTTGGATTCATATGAGCCATGCTTACTGTTCTAAGGTCTTAGCTGGACCTGGGgcattatagatcccaagcagttcttttataagaactgagaaataccctttacttagcaggtggcggcagaaggatcagaagttcaagggcatccttggctacttagacagtttgagactaccctgcagccatggaaatggagggaaggaaggagggatggaggaagagggaggacacattttaccatctcatttgctttcttctttcttgccctctccaactgggctgctatttgactgcagatcgtagtaccaccactgcctgggaaagccttgaagcggcagctccctttccgaggagatgaagggatctttgaggagtctttcattgaagaaaggaggcaaggcctcgaacagtttattaacaagtaagccaaattcttggctgctttgcatactgcttttgccattatgaaatgtttctttaccaccctcccttccccatttttctctctcctccctgtcctcctTCATTCCATACTTGGTTCTTTCCCTTTTTGCCTGGGAATGACTCCACAGCCTTGCATGTGCTAGTCAAGCACTCTACCTCTAAACTATTATACTCCCAGCCCATCATGATGTTCTTATATATATTTTTGTCTACTTTG

This mutation is a 469 bp deletion beginning at Chromosome X position 101,214,299 bp and ending after 101,214,767 bp (GRCm38/mm10).

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
50451 GAA ATG GAG GGA AGG AAG GA Wild type Forward A
50452 AAT AGC AGC CCA GTT GGA GA Wild type Reverse A
50453 CTC TGT CCT TCC ACA GAA ATC A Mutant Forward A
50454 AAG GGA AAG AAC CAA GTA TGG A Mutant Reverse A
50455 Fluorophore-1 AGG AAG AGG GAG GAC ACA TTT TAC C Quencher-1 WT Probe
50456 Fluorophore-2 AAG GTC TTA GCT GGA CCT GGG TC Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
50451 0.40 uM
50452 0.40 uM
50453 0.40 uM
50454 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.