Stock Number:001927
Strain Name:C57BL/6-Tg(APOA1)1Rub/J
Tg(APOA1)1Rub
Tg(APOA1)1Rub
Separated QPCR
3.1
22-July -2008 24-September-2009
Notes: Taqman qPCR protocols are run on an ABI 7500, 7700, 7900 or the Roche LightCycler 480. Use an appropriate instrument-specific Fluorophore/Quencher combination. The transgene genotype is determined by comparing δCt values of each unknown sample against known homozygous and hemizygous controls, using appropriate endogenous references.
Protocol Primers 
Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type
oIMR1225 - TGT CTC ACC CTC CAG CCT AAA - Transgene
oIMR1226 - TGT TCG CGC AGC TTG CT - Transgene
oIMR1544 - CAC GTG GGC TCC AGC ATT - Internal Positive Control Forward
oIMR3580 - TCA CCA GTC ATT TCT GCC TTT G - Internal Positive Control Reverse
TmoIMR0009 6-FAM AAC TGG GAC AGC GTG ACC TCC ACC TT TAMRA Transgene
TmoIMR0107 JOE NHS Ester CCA ATG GTC GGG CAC TGC TCA A Black Hole Quencher 1 Probe
Reaction/Components 
Reaction Component Volume (µl) Final
Concentration		 Total Volume (µl)
2 X Universal Master Mix 10.00 0.80 X 10.00
40 uM oIMR1225 0.25 0.40 uM 0.25
40 uM oIMR1226 0.25 0.40 uM 0.25
10 uM TmoIMR0009 0.38 0.15 uM 0.38
ddH2O 4.13 - 4.13
DNA 10.00 - 10.00
Cycling 
Step # Temp °C Time Note
1 50 2 min -
2 95 10 min -
3 95 15 sec -
4 60 1 min repeat steps 3-4 for 40 cycles
Reaction/Components 
Reaction Component Volume (µl) Final
Concentration		 Total Volume (µl)
2 X Universal Master Mix 10.00 0.80 X 10.00
40 uM oIMR1544 0.25 0.40 uM 0.25
40 uM oIMR3580 0.25 0.40 uM 0.25
10 uM TmoIMR0107 0.38 0.15 uM 0.38
ddH2O 4.13 - 4.13
DNA 10.00 - 10.00
This genotyping protocol is used for the following strains:
 
Stock Number Strain Name
001927 C57BL/6-Tg(APOA1)1Rub/J
Genotyping resources and troubleshooting
This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.
Cycling 
Step # Temp °C Time Note
1 50 2 min -
2 95 10 min -
3 95 15 sec -
4 60 1 min repeat steps 3-4 for 40 cycles