Stock Number:017434
Strain Name:FVB;B6-Cramp1lTgTn(sb-rtTA,Tyr)2447AOve/J
Cramp1lTgTn(sb-rtTA,Tyr)2447AOve
Cramp1lTgTn(sb-rtTA,Tyr)2447AOve-3'
Separated PCR
1.0
12-October -2011 16-November -2011
Notes: This assay will NOT distinguish hemizygous from homozygous transgenic animals.
Expected Results:
Transgene = 550 bp
Internal Control = 324 bp
Gel Image 
Image Image file
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Protocol Primers 
Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type
13134 - CGT CTG TTG TGT GAC TCT GGT AAC - Transgene Reverse
13292 - GGA CAC ATA CTT AAC ATC CCA CAA - Transgene Forward
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)
10 mM dNTPS-kapa 0.24 0.20 mM 0.24
25 mM MgCl2 0.96 2.00 mM 0.96
20 uM 13292 0.30 0.50 uM 0.30
DNA 2.00 - 2.00
20 uM 13134 0.30 0.50 uM 0.30
5 X Kapa 2G HS buffer 2.40 1.00 X 2.40
ddH2O 4.55 - 4.55
5 mM 10x Loading Dye 1.20 0.50 mM 1.20
2.5 ul Kapa 2G HS taq polymerase 0.05 0.01 ul 0.05
Cycling 
Step # Temp °C Time Note
1 94 2 min -
2 94 20sec -
3 65 15sec -1.5 C per cycle
4 68 15sec repeat step 2-4 for 10 cycles
5 94 15sec -
6 50 15sec -
7 72 10sec repeat steps 5-7 for 25 cycles
8 72 2 min -
9 10 10 Forever
Reaction/Components 
Reaction Component Volume (µl) Final
Concentration		 Total Volume (µl)
ddH2O 4.55 - 4.55
5 X Kapa 2G HS buffer 2.40 1.00 X 2.40
25 mM MgCl2 0.96 2.00 mM 0.96
10 mM dNTPS-kapa 0.24 0.20 mM 0.24
20 uM oIMR7338 0.30 0.50 uM 0.30
20 uM oIMR7339 0.30 0.50 uM 0.30
5 mM 10x Loading Dye 1.20 0.50 mM 1.20
2.5 ul Kapa 2G HS taq polymerase 0.05 0.01 ul 0.05
DNA 2.00 - 2.00
This genotyping protocol is used for the following strains:
 
Stock Number Strain Name
017434 FVB;B6-Cramp1lTgTn(sb-rtTA,Tyr)2447AOve/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 94 2 min -
2 94 20sec -
3 65 15sec -1.5 C per cycle
4 68 15sec repeat step 2-4 for 10 cycles
5 94 15sec -
6 50 15sec -
7 72 10sec repeat steps 5-7 for 25 cycles
8 72 2 min -
9 10 10 Forever