Protocol 35929: Probe Assay - Igf2bp1<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

>chr11:95974915-95975059 145bp GTCCTGCACTGTCTGTGTTGA AGAAATCAGGGTTGCCCTTG

Mut= 145 bp

Wt= 145 bp

Fam=Mut

Hex=Wt

Sequence

Wt Sequence (deletion in lower case):

 AGCATGGTGGCCTGATGTGTCCCATGTCCACTGTCAAGAAGTTTAGAATATTTGAGCCTGGAATTCCTTCTCTGGGAAAGTGTGTGATATCCAAGCTTTACAGAATGGATTTAGATAATTCCAAATCTCTTGAGCCATTGAGAGAAGGAAACCATTTTTTTCTTGACATATTACCTCTGTAAATGGCTAATCCCACccactgggtgttgcttgtttgctaattgaactttcccagggtaagtttgcttccagtgtttttgtgtgtgtgtgtttgtgtgtgtgtgtgtgtttttgtgtgtgtgtgtgtgtgtgtgtgagggaggtcatctgtgaattgttttttgcttgtcctttgagtctgttcccagcgtggtagtctgctgcttatcctgttaacctacagacaggttggggtttgtcagcagagctctaactctggtggactattctgttgtctccacagtgaacactgaaagtgagacagctgtggtcaacgtcacctactctaaccgggagcagaccaggcagtgagtacctcagaagcggggctgggggctgttctgtgggtgggtccaggcttctgatattggttcagctagccctaaggtcccacaaaaggtcttttaggggtatgtttcagaaatcagaaaattctcagtaaaaatgatctgcaggaagggggatgactgggagaagagatggcttttgggtccctgggaacaacagagtcctgcactgtctgtgttgatggtgccatcttttggttatgagggggaaactgcctcgttcaaaacTGCATATATTTCTTTTTTTGAAATATATAGTGGCCTCTAATTTATTTGCCTTGGTAGCCAAGGGCAACCCTGATTTCTACCCCATTTTCAGAGCATTGTGATTGTGGGTGTGGGTTACTGTGCCTATATCAGGTATGATGCTGGATGGTTTAAGAGCCTGGGTGATGGGTTTCTAGGTCCTTTGAGTCCCCACCACCACCACAGG

This mutation is a 598 bp deletion beginning at Chromosome 11 position 95,974,993 bp and ending after 95,975,590 bp (GRCm38/mm10).

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
47340 GTC CTG CAC TGT CTG TGT TGA Wild type Forward A
47341 AGA AAT CAG GGT TGC CCT TG Common A
47342 TTG AGC CAT TGA GAG AAG GA Mutant Forward A
47343 Fluorophore-1 ATC TTT TGG TTA TGA GGG GGA AA Quencher-1 WT Probe
47344 Fluorophore-2 CTG TAA ATG GCT AAT CCC ACT GC Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
47340 0.40 uM
47341 0.40 uM
47342 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.