Protocol 33701: Probe Assay - Dusp23<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

>chr1:172632559-172632661 103bp CGACCAATTTGTGAAGATCG GCCCACTCATTTCCTTCCAG

Mut= 91 bp

Wt= 103 bp

Fam=Mut

Hex=Wt

Sequence

Wt Sequence (deletion in lower case):

TACGTGGCCCGCTAGGGCAAGTGCTGAGCGGGGACGGAATCCAGCCCCAGAGGGGGGGTGACCCAATCTCCCGGCAGAGTAGGAAAGGCCAGCCTCGCCCTGAGTAAACTCCCCCCACGATGGgcgtgcaaccccccaacttctcctgggtgcttccgggacggctggccggactggcgttgccccggctgcccgcgcactaccagttcctgctggaccagggtgtgcggcacctggtgtccctgacggagcgcggaccccctcacagtgacagctgtcccggcctcacgctgcaccgaatgcgcatccctgacttttgcccgccgtccccggaacagatcgaccaatttgtgaagatcgtggacgaggccaatgcccggggagagGTCAGCACGCTCGTGAGCCAGAGCGAGAGGGCGGGGCCTGGAAGGAAATGAGTGGGCGGGGCCGGAGCTGGAGTCCGAAAAGAGTGGGAGAGCCTGGGGTTGGGGGCGGGGGTGACAGGGCTAAGTGGGGTGGGAG

This mutation is a 263 bp deletion beginning at Chromosome 1 position 172,632,616 bp and ending after 172,632,878 bp (GRCm38/mm10).

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
42007 CGA CCA ATT TGT GAA GAT CG Wild type Forward A
42008 GCC CAC TCA TTT CCT TCC AG Common A
42009 CAG CCT CGC CCT GAG TAA AC Mutant Forward A
42010 Fluorophore-1 ACG AGG CCA ATG CCC Quencher-1 WT Probe
42011 Fluorophore-2 CGA TGG GTC AGC ACG C Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
42007 0.40 uM
42008 0.40 uM
42009 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.