Tg(SMN2*A111G)591Ahmb
Tg(SMN2*delta7)4299Ahmb
QPCR
7.0
13-March -2012 13-March -2012
Notes: Taqman qPCR protocols are run on an ABI 7500, 7700, 7900 or the Roche LightCycler 480. Use an appropriate instrument-specific Fuorophore/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
13739 6-FAM CTT CTG GAC CAC CAA TAA TTC CCC CAC C

Zen probe with Iowa Black

 Tg Probe
oIMR1544 - CAC GTG GGC TCC AGC ATT - Internal Positive Control Forward
oIMR3580 - TCA CCA GTC ATT TCT GCC TTT G - Internal Positive Control Reverse
oIMR3675 - TGC TGG CTG CCT CCA TTT - Transgene
oIMR3676 - GCA TCA TCA AGA GAA TCT GGA CAT - Transgene
TmoIMR0105 Cy5 CCA ATG GTC GGG CAC TGC TCA A Black Hole Quencher 2 IC Probe
Reaction/Components 
Reaction Component Volume (µl) Final
Concentration		 Total Volume (µl)
2 X Kapa Probe Fast QPCR 5.72 0.95 X 5.72
40 uM oIMR3675 0.13 0.43 uM 0.13
40 uM oIMR3676 0.13 0.43 uM 0.13
40 uM oIMR1544 0.13 0.43 uM 0.13
40 uM oIMR3580 0.13 0.43 uM 0.13
5 uM 13739 0.38 0.16 uM 0.38
5 uM TmoIMR0105 0.38 0.16 uM 0.38
DNA 5.00 - 5.00
Cycling 
Step # Temp °C Time Note
1 95 10min -
2 95 15sec -
3 60 1min repeat steps 2-3 for 40 cycles
This genotyping protocol is used for the following strains:
 
Stock Number Strain Name
005025 FVB.Cg-Tg(SMN2*delta7)4299Ahmb Tg(SMN2)89Ahmb Smn1tm1Msd/J
005026 FVB.Cg-Tg(SMN2)89Ahmb Tg(SMN1*A2G)2023Ahmb Smn1tm1Msd/J
007951 STOCK Smn1tm3(SMN2/Smn1)Mrph Tg(SMN2*delta7)4299Ahmb Tg(SMN2)89Ahmb/J
008203 STOCK Smn1tm1Msd Tg(ACTA1-SMN)63Ahmb Tg(SMN2)89Ahmb/J
008209 FVB.Cg-Smn1tm1Msd Tg(ACTA1-SMN)69Ahmb Tg(SMN2)89Ahmb/J
008212 STOCK Smn1tm1Msd Tg(Prnp-SMN)92Ahmb Tg(SMN2)89Ahmb/J
008604 FVB.129(B6)-Smn1tm5(Smn1/SMN2)Mrph/J
008782 FVB.Cg-Smn1tm1Msd Tg(SMN2)89Ahmb Tg(SMN2*A111G)588Ahmb/J
008783 STOCK Smn1tm3(SMN2/Smn1)Mrph Tg(SMN2*delta7)4299Ahmb Tg(SMN2)89Ahmb Tg(CAG-cre/Esr1*)5Amc/J
009134 FVB.Cg-Smn1tm1Msd Tg(SMN2)89Ahmb Tg(SMN2*A111G)591Ahmb/J
017596 STOCK Gt(ROSA)26Sortm1.1(rtTA,EGFP)Nagy Smn1tm1Msd Tg(SMN2)89Ahmb Tg(SMN2*delta7)4299Ahmb Tg(tetO-SMN2,-luc)#aAhmb/J
Genotyping resources and troubleshooting
This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.