Protocol 35405: Probe Assay - Serinc5<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

>chr13:92683100+92683210 111bp CAGGGAGTTTATTGTTTCTACTAGGG TTTCCCATAAGGAGGCACAC

Mutant=  100 bp

Wild Type = 111 bp

Sequence

GTCCAGCTAGAAACTCTAGCGGATAGGTTGTTCTTATTCAGTCTTGGTGCTGATTTGCACATGGCTGTTTTTACCATGCATTTGACTAAGACTGAACACTCACAGGCAGCTCCTCTCagcctagttctaacaggcaacttttgcatcttctctctcagattcccttttttgaagagttctgtaaaaagactcaagccggtggtgacgcctgtgagaacctggtagggtattctgcggtgtacagagtctgctttggaatggcttgtttctttgctctgttttgcctgctgaccttaaaagtcaacaacagcaaaagttgccgggcctacattcacaacgggtaagtctcttgttcttccaagggtgtcctagggtctgctcgtcctcactgactgacgttctgctagttgacatttgacactgatgtggggaggatgggctgtgtgtcactggtagcctaagagcggcagtgcctggtgccataataagaaattcagtatacacagggagtttattgtttctactagggtcttcccctgagtcatttccatcctaccgTGAAGAAGAGTTTTATACAATCAGAGGCCATTCCAAGTGTGCCTCCTTATGGGAAAAGCCACTCTGTGGCACTCCATCACTTCCCCTTGATGTGGGAAGAACACTACCAACAACAGAAACATGAGCTAGCAGGCGGCCTTACAAGGA

 

A 441 bp deletion beginning at Chromosome 13 position 92,682,714 bp and ending after 92,683,154 bp (GRCm38/mm10).

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
45979 CAG GGA GTT TAT TGT TTC TAC TAG GG Wild type Forward A
45980 TTT CCC ATA AGG AGG CAC AC Common A
45981 CCA TGC ATT TGA CTA AGA CTG AAC Mutant Forward A
45982 Fluorophore-1 TCC CCT GAG TCA TTT CCA TC Quencher-1 WT Probe
45983 Fluorophore-2 AGG CAG CTC CTC TCT GAA GAA G Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
45979 0.40 uM
45980 0.40 uM
45981 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.