Protocol 35238: Probe Assay - Exoc3l2<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

>chr7:19474494+19474610 117bp TTGCACTGGCACAATCATGT GCTACATGAAACCCAATCCCTA

 

Mut= 123 bp

Wt= 117 bp

Fam=Mut

Hex=Wt

 

Sequence

Wt Sequence (deletion in lower case):

 TGCACGTATATGTGGAGGCCAGAGGAAACCTTGCCCGAATCAGTTCTACCATGTGGGCCCTGGGAATGGGGAAGAAACTCGTTAAACTTGGATGGTCGTTAAACTTGACAGCATGCACGTTTGTCCACTCCATCTCATTTGCCTGTTTGTTTGTTGTGACTTGTCCTCAACTCTGTAGCCCACTctcatgagtgcaattgctgaggtcaaaggcaagcgtcaccctcactctcatgttggggggaactcattttgggtggcaagccctgactctctcctcatcccccgcttctcactttgcgggtactgggtatgtgccccgcttcctgacaccttttctctccccaccctgcacagtgctggagatcctgggcctgatccagaggcgcgagctggcgcgagcggacgagcacatcctggagctggaggcggaggcgccgacggcggaggccgaggcgggcgcggggcggcgcgcgcgggacgtggcgctgctgtacgaggcgctgcagcgcgagctctgggcgctggtgcgcgagtcgctggcttccccgggacccggcaccggcgcggtgtcgcagctgggcctcgtgctgctgcaggaggaggcggcggacgcgcgccgcggccccggggcggcccgcaagctgcgcacgcactgggccgaggcggtggcgcgcgcagcgcgcgaacgtctggaggaggcggcggcgccgggcgcgcgcggggcgctggctgggcagctggaggcgctgcgcgcgcgcctgttggaggacgtggccgctgtgcgcggacgcctggcgcccgcctaccctgcggggctgcgcgccggcgccgtctacctgcgtggctaccacgaggcactggccgagtggctcggggcggccgcgcgccgcaggctcccgctggcggatcgttatgcgctgttgcactggcacaatcatgtgtaccccaggtgagcccgggtcgccccggGACGCGCTCACCTTAACCCCAGGCACCTTGTTCAAGATACTGGATTTAGGGATTGGGTTTCATGTAGCCCAGGTTGACTTGAGGTTTTCTGAGACAGGGTCTTGCTCTGTAGACTAGGCTTGCCCTAAACTCAGAGATCCGCTCGCTTTTACCTCTTGAGAGCTCCTATAAAAGCGAGCACCGCTGCACCC

This mutation is a 788 bp deletion beginning at Chromosome 7 position 19,473,755 bp and ending after 19,474,542 bp (GRCm38/mm10).

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
45591 TTG CAC TGG CAC AAT CAT GT Wild type Forward A
45592 GCT ACA TGA AAC CCA ATC CCT A Common A
45593 CCA TCT CAT TTG CCT GTT TG Mutant Forward A
45594 Fluorophore-1 CAG GTG AGC CCG GGT C Quencher-1 WT Probe
45595 Fluorophore-2 TAG CCC ACT GAC GCG C Quencher-2 MUT Probe

Reaction A

Component Final Concentration
ddH2O
Kapa 2G HS buffer 1.30 X
MgCl2 2.60 mM
dNTP KAPA 0.26 mM
45591 0.50 uM
45592 0.50 uM
45593 0.50 uM
Glycerol 6.50 %
Dye 1.00 X
Kapa 2G HS taq polym 0.03 U/ul
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.