Stock No: 033278
Protocol 36434: Probe Assay - Pex7<em4Lutzy>
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

Mutant=  108 bp

Wild Type = 99 bp 

>chr10:19904880-19904978 99bp GTCAGCATGTCTTTGGTGGA GGAAGCACAGCCCAGGAA

Sequence

Wt Sequence (with deletion in lowercase):
 
TCTCACAAGTGCATGCTGTGAGTCACAGGTGCACTACAATAAAAGCAGGTGATGTCAGCATGTCTTTGGTGGATACCTCATATGAGTGTTTGGAAACCTCCTTTCAgtttaactttggagctatctacctgcccttcctgggctgtgcttccaggccactgtggagtacctgctctatttatctcttgtagttttcaagatcatctgaggtaatgtgtgttacaatatgtgaaaaacaaggaaaaaaacagtaactataaaattaagatgttaatcattttgtgatctgcagtgcatgtattttcttttctggcgtagctttgactggaatgacggcttgtttgacgttacctggagtgaaaacaatgagcatgtccttgtcacctgtagtggtgatggctcactgcagctctgggacactgccaaagccacaggccctctccaagtctataaagagcacacgcaggaggtatgatgatggcttcccttattgtttgtttgaaatctctttgggatgaacacagggaaatgctttcaggggacaaggttaagctttaatgtttcttattcatagtagatgtcacttattattctgttattcaaacataaagtacttcttagcctaagacaagacaagaaaaggtcaggcttctcattaggcaacagcagtgcctctagcaggcacgGTGGCATCCTTCCCTCCAGAGGGCCTCAGCCTTCCTTATGCCCTATGAACACGCCTACACGCAGTGCCTCAGGAGAGGGTGGGCGGGCTGTTCGCAGGCTCTCTCACGAT

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
48755 GTC AGC ATG TCT TTG GTG GA Common A
48757 GGA AGC ACA GCC CAG GAA Wild type Reverse A
48758 Fluorophore-1 ACT TTG GAG CTA TCT ACC TGC C Quencher-1 WT Probe
48759 GGC GTG TTC ATA GGG CAT AA Mutant Reverse A
48760 Fluorophore-2 CCT CCT TTC AGT GGC ATC CT Quencher-2 MUT Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
48755 0.40 uM
48757 0.40 uM
48759 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.