Protocol 33836: Probe Assay - Cabp2<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

>chr19:4084768+4084882 115bp GGCCTTGGGTATGTTGTGAG CGATCCTGCAAAGTACACAGA

Mut= 109 bp

Wt= 115 bp

Fam=Mut

Hex=Wt

Sequence

Wt Sequence (deletion in lower case):

 

GTCCTGGGGTCCCATGGCAGGTCACAGGGAAAGGGATCCCTAAGACACCCAGACAGATGACCAGGACCGGGCAGAATCAGGCACAGGCCTTGGGTATGTTGTGAGGTGGGCTGGATGAGAAACCTCAAGACACAGAAAGAGGGTgccgaggtgtgggcaggccaaggaactcctaactatctgtgtactttgcaggatcgagagctgagaccagaagagatcgaaggtaatgggttcagggagtgggtagttgaggtggagtgtggggtgggggttacaagaagaggtttggggctagggcctctgcaaggtccaggaggagcgagaaggacatggggtctgggcagggagggctggcaaaacctcccttctttgccaaccccttggtctcagtgaccacactggtcaaggactgcttttatatttgtccatctggctgacacttcctggctgtccactgtgggcctggccccttgctaaccctgctaaccaggtagattaagggccccatcttctggagatagctacagctgcagctgcatagagcactcttaggaaaaccaatgactgggctgactaggaagaaatgggcagggaagtcttcctggtagcttttctgtctcttctcccctcccaccttctcttccttctctttttaatcacctagaacagagatgggcacatgacagatgcttcttaagcattggatgagagaacaaaattttaacactacacatgcaaaggctcttgggtaacaactaggaactgaggtttaacaacaagaagggactggagcaaagggaaaagatggaccggccccaaccggcagccaggaaggggcgctcagaaagggactaacctcactctgtccggcagagctgcagatcgccttccaggagtttgaccgagaccgggacggctacatcggctaccgggagctgggtgcctgcatgcggacactgggctacatgcccacggagatggagctcattgagatatcacaacagatcagtatgtgcttcagacccaacagagcctcatgccccagccctggcccagccctgaggtagacgtgtgcgctgttccaaacgcccaggaTATAAAGTTGTTATTAAAGGGAGAGCAGGGAGGATGGCTCCAGTAAAGAGACTGAAGCTCAGAGAGGTGAAGTATTGTCTAGAGTCACACAGCAGGGGCTCAGAGAAACCAAGAAACTGTGACTTAAGATCCTAGTA

 

This mutation is a 954 bp deletion beginning at Chromosome 19 position 4,084,827 bp and ending after 4,085,780 bp (GRCm38/mm10).

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
42297 GGC CTT GGG TAT GTT GTG AG Common A
42298 CGA TCC TGC AAA GTA CAC AGA Wild type Reverse A
42299 CTC TTT ACT GGA GCC ATC CTC Mutant Reverse A
42300 Fluorophore-1 CAG GCC AAG GAA CTC CTA ACT Quencher-1 WT Probe
42301 Fluorophore-2 AGA GGG TTA TAA AGT TGT TAT TAA AGG GA Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
42297 0.40 uM
42298 0.40 uM
42299 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.