Protocol 35991: Probe Assay - Lurap1l<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

>chr4:80910531+80910642 112bp AGGCAGGATTTGCATTTCAG ACTGGCAGGTGGACCAAAG

Mut= 114 bp

Wt= 112 bp

Fam=Mut

Hex=Wt

Sequence

Wt Sequence (deletion in lower case):

 TAAAATTTAGCCATTCTTCTGCCTCTAATGTTAAAAATGTGTTCTACAGGTTGTAGCTGTCAGGGGACTTCAGCAGGGCAACTTTTAACTTAGTAACCTAATATGGGTATGCTCAGCTGAGGCAGGATTTGCATTTCAGTTTCTTAACATCAGGGTTGGGAAGATTTTGCTGCCCTTcttgtctgtgacatacggtacaggggttggctctgctttggtccacctgccagtcgagtttaaagtttccgaggctcggaggaacacaatgacttggatgagaccgactaaaaggaaaggacactgctgcagcaaccaggaagcccttgaaccccagctaagctaaccgtgtctttttcttttttctttctaaaatgcgcctacaccgcctacacacaggacgaacccacttctttcattgatcagatttttttttttttttcctgaccgtctaagatagtctgatccttggcgacatatggctggaaaatgaaacgacccgggagtgtggccacctcgttattctgtgtggtgtaataccagatgggcaccgtgaaccgcgctgggatgtgaatggacagtttgaaatgggaatttccccgcatcgagcaggttccatttctctcttgggtctctcggtataacgtactagcttttgcccttcttctctgcagtatcaattaactcaacttggcggtggagtggccagctaggaaaggaatgaagcgatctggctaatcttgctggctgtggagcggcagagcctgaaactgcagccgggtgcagatctgccgactgcgcccggggcatggagcgcacgggcgaccgcggggactcaggcccggccagaggagcccagtactgagctccggagtccgccaggccgccgccgccgcctcctcgcgccctctgcctttactgtcagtgtcgcccctcgcaaagagtcatggaagacagcccgctgccagacctcagagacatcgagctgaagctggggcgcaaagtgcccgagagcctagcgcgctcgctgcgcggggaggagctggcgcctcgggaaggggctgcggaccccagcggggttggtgggagctgcagcagcagcagtagctgcagcagctttgcgccctccgtgtcctcctcgtcgtcatcctcccctgcctctggctctccacgacgtagccaccccagcgccctggagaggttagagaccaaactgcacatcctcagacaggagatggtgagtgtgatgcgtcagaggcgacttggggacccggggcctgggacccgagagggtggcacaggagcggggggggggggggggggagaagctgggaggaggtgtgcgggagcgcgcctctggactcggctacagctaagcttgaaccctgaggactttgctcggtcacggggtgccctggcagctcattcggctctactttctgatagagctactccctgcgcaccactctgcagctgccagccgcggattctcccgcactagcCTGCTAAAGGCCGGCAGTTACTGCGCGGATCTGGGCCTGCTGTGAGCCGTTTATGTTTTGCCTTGGTTCAAAGGGTGGTTTAGTCCAATGCACTTAGCGCAGAAATGCCTCCCAGCTGGCCGGGAGGTGGCGCGGCGGGGTCCTTGCAGCCTGCTGTAGTTTG

This muatation is a 1322 bp deletion beginning at Chromosome 4 position 80,910,589 bp and ending after 80,911,910 bp (GRCm38/mm10).

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
47508 AGG CAG GAT TTG CAT TTC AG Common A
47509 ACT GGC AGG TGG ACC AAA G Wild type Reverse A
47510 ACA TAA ACG GCT CAC AGC AG Mutant Reverse A
47511 Fluorophore-1 ATA CGG TAC AGG GGT TGG CT Quencher-1 WT Probe
47512 Fluorophore-2 CTG CCC TTC TGC TAA AGG C Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
47508 0.40 uM
47509 0.40 uM
47510 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.