Stock No: 008027
Protocol 22218: QPCR Assay - bpck QPCR
Version 4.0

Notes

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

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
8009 Fluorophore-1 TCA AAA ATG CTC ATA GCC AGT CTG CGA A Quencher-1 Tg Probe
8012 CCA TTG CCT ACA GTG ACA TTG AC Transgene Forward A
8013 GGG CAC TGA ATA ATG AAG AAC TCA Transgene Reverse A
oIMR1544 CAC GTG GGC TCC AGC ATT Internal Positive Control Forward A
oIMR3580 TCA CCA GTC ATT TCT GCC TTT G Internal Positive Control Reverse A
TmoIMR0107 Fluorophore-2 CCA ATG GTC GGG CAC TGC TCA A Quencher-2 IC Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
8012 0.40 uM
8013 0.40 uM
oIMR1544 0.40 uM
oIMR3580 0.40 uM
Tg Probe 0.15 uM
IC Probe 0.15 uM
DNA

Cycling

Step Temp °C Time Note
1 95.0 --
2 95.0 --
3 60.0 -- repeat steps 2-3 for 40 cycles
JAX uses a very high speed Taq (~1000 bp/sec), use cycling times recommended for your reagents.

Strains Using This Protocol

Stock Number Strain Name
008044 B6C3Fe a/a-bpck/J
008027 C57BL/6J-Tg(RP24-229O13)53Dn
008028 C57BL/6J-Tg(RP24-229O13)54Dn/J
3 strains use this protocol