Protocol 36482: Probe Assay - Armc3<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

>chr2:19235367+19235516 150bp ACAGAGCAGTGACCACATCC GCTGCCTTTTTGCTTTCA

Mut= 147 bp

Wt= 150 bp

Fam=Mut

Hex=Wt

Sequence

Wt Sequence (deletions in lower case):

 TTCAAGGCCAATCTGGACAACACTGTGGAATCTGAGCCCCCTTCCCTCCCACGATGTTAATGCATCAAATAAAAATAATTAAAGATGCAAAATATAAGAGGTATATGTGACCCATATCAGGAAAACAATCAATAATCAATTAGAGGAAATAAAATAGGAAGCTAAAAAAATTCAGTTACCAAAAAGAGAAGACAGAAAAAAGAGAGAGTAGAGACAGATAGCAAGATGGTCAATAGATATCAACCATATTAATAATCATACTAGATTATTGGTTTCCTTTTTAAACAGGAACTTTGCAGCATACTTACAGAGCAGTGACCACATCCTTTTGTAAGACTGCAGCGTCTTTAAATTTTAAGccactgttaattttgatgtgacagttacatgaacctaattacttttcctttttctccttcagtttgacccaatcacaattgaaagcaaaaaggcagccactgtggtgctgatgctcaagtctccagaggaagacatcttagccaaagcctgcgaagccatctataagtttgctctgaaaggtttgttcttttggtgttaaactttctatctcgtttctgtatagtactatatatatataataatatacataccaggtgctctgataagaataaatgccacagaagcaaaaagagcagttagataaacttgcattggatcatacccaaagtataagacaaatatccatgtgtacaatgttgatataagagttaaactgaaggcggccccaccccttcctccctgctcCCTTCCCCACTGGCCcctttcACTCAGGGTATTCTCCATTTCCACTTTGTTCTTCTACCCCCTCCTTTTCCCTCCCAGCCTCCTTTGCCCAGGCTCATTGGCCCTTCCTAATTTCCTCGGCTTTACCGCCCATTTTCATGCTAATTTTGTAATTTCATCTTCCTTTACAGCTGAATAAAATTCCATTCAAATAAAATTTAAATGATCAACAGGGACAGATAGTTGTATTCAATTAAAAACCAAAACCAAAACCAAACTGTATGCTGCCTTGAAGAAAGATCTTTAGAATATTAAGGCGCAAATAAAAGGTAAAAAGATATCTCATGAAAGGACTAATCAATATAAACATGGAGTGATTATATTAA

This mutation is a 406 bp deletion beginning at Chromosome 2 position 19,235,420 bp and ending after 19,235,825 bp (GRCm38/mm10). In addition, there is a 6 bp deletion (CCTTTC) 15 bp downstream the 406 bp deletion.  

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
48856 ACA GAG CAG TGA CCA CAT CC Common A
48857 GCT GCC TTT TTG CTT TCA Wild type Reverse A
48858 CAA TGA GCC TGG GCA AAG Mutant Reverse A
48859 Fluorophore-1 CTC CTT CAG TTT GAC CCA ATC A Quencher-1 WT Probe
48860 Fluorophore-2 ATT TTA AGC CTT CCC CAC TGG Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
48856 0.40 uM
48857 0.40 uM
48858 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.