Genotyping Protocols Database
Stock No: 012466
Protocol 30002: QPCR Assay - Lrrk2/LRRK2
Version 3.0

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
11569 CTC ATG GAA GAA TGG TTT CC Transgene Forward A
11570 GAT GTT CTT CAC CAT CAT TAA AAC T Transgene Reverse A
11571 Fluorophore-1 AGG AGA AAC TCT GTT GAA GAA ATG GGC A Quencher-1 Tg Probe
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
TmoIMR0105 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
11569 0.40 uM
11570 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
012466 B6.Cg-Tg(Lrrk2)6Yue/J
012467 B6.Cg-Tg(Lrrk2*G2019S)2Yue/J
2 strains use this protocol