Stock Number:013725
Strain Name:B6;SJL-Tg(LRRK2)66Mjff/J
Tg(LRRK2)66Youy
Tg(LRRK2)
Melt Curve Analysis
3.0
24-April -2011 26-June -2012
Notes:

This assay will NOT distinguish hemizygous from homozygous transgenic animals.

Melting curve analysis is done using a Roche Light Cycler 480.
Expected Results: Transgene = 154 bp
Internal Positive Control = 324 bp
Gel Image 
Protocol Primers 
Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type
8752 - TGA TTC TCG TTG GCA CAC AT - Transgene Forward
8753 - GCC AAA GCA TCA GAT TCC TC - Transgene Reverse
oIMR7338 - CTA GGC CAC AGA ATT GAA AGA TCT - Internal Positive Control Forward
oIMR7339 - GTA GGT GGA AAT TCT AGC ATC ATC C - Internal Positive Control Reverse
Reaction/Components 
Reaction Component Volume (µl) Final
Concentration		 Total Volume (µl)
ddH2O 1.60 - 1.60
2 X SYBR Green Master Mix (Roche) 6.00 1.00 X 6.00
20 uM 8752 0.60 1.00 uM 0.60
20 uM 8753 0.60 1.00 uM 0.60
20 uM oIMR7338 0.60 1.00 uM 0.60
20 uM oIMR7339 0.60 1.00 uM 0.60
DNA 2.00 - 2.00
Cycling 
Step # Temp °C Time Note
1 94 5 min -
2 94 30sec -
3 68 1 min -
4 72 1 min -
5 - - Go to step 2-4, 35 times
6 72 2 min -
7 10 - hold
This genotyping protocol is used for the following strains:
 
Stock Number Strain Name
009604 FVB/N-Tg(LRRK2*R1441G)135Cjli/J
009610 FVB/N-Tg(LRRK2)1Cjli/J
012445 B6.FVB-Tg(LRRK2)WT1Mjfa/J
012446 B6.FVB-Tg(LRRK2*G2019S)1Mjfa/J
013583 B6.Cg-Tg(tetO-LRRK2)C7874Cai/J
013725 B6;SJL-Tg(LRRK2)66Mjff/J
021913 B6.FVB-Tg(LRRK2*Y1699C)C7.11HMjfa/J
370469 FVB/N-Tg(LRRK2*R1441G)135Cjli/J
370470 FVB/N-Tg(LRRK2*G2019S)1Cjli/J
370471 FVB/N-Tg(LRRK2)1Cjli/J
370472 B6;SJL-Tg(LRRK2)1Youy/J
370473 B6;SJL-Tg(LRRK2*G2019S)1Youy/J
370474 B6;SJL-Tg(LRRK2*R1441G)1Youy/J
Genotyping resources and troubleshooting
This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.