Stock Number:017800
Strain Name:B6.Cg-Fig4plt1/MmJ
Fig4plt1
Fig4plt1
Separated PCR
1.0
05-April -2012 18-April -2012
Expected Results:

Mutant = 390 bp
Heterozygote = 333 bp and 390 bp
Wild type = 333 bp
Gel Image 
Image Image file
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Protocol Primers 
Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type
14432 - CTG GCT TCT TTG GTG ACA GG - Common
14433 - TCG TGT TTG CTG GAA AGT GA - Wild type Reverse
14434 - GGA GCT AAG GCA ATT TCA TAC TG - Mutant Reverse
Reaction/Components 
Reaction Component Volume (µl) Final
Concentration		 Total Volume (µl)
20 uM 14433 0.30 0.50 uM 0.30
2.5 ul Kapa 2G HS taq polymerase 0.05 0.01 ul 0.05
25 mM MgCl2 0.96 2.00 mM 0.96
10 mM dNTPS-kapa 0.24 0.20 mM 0.24
20 uM 14432 0.30 0.50 uM 0.30
5 mM 10x Loading Dye 1.20 0.50 mM 1.20
5 X Kapa 2G HS buffer 2.40 1.00 X 2.40
ddH2O 4.55 - 4.55
DNA 2.00 - 2.00
Cycling 
Step # Temp °C Time Note
1 94 2 min -
2 94 20sec -
3 65 15sec -1.5 C per cycle
4 68 10sec -
5 - - Go to step 2-4, 10 times
6 94 15sec -
7 50 15sec -
8 72 10sec -
9 - - Go to step 6-8, 28 times
10 72 2 min -
11 10 - hold
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 14432 0.30 0.50 uM 0.30
20 uM 14434 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
017800 B6.Cg-Fig4plt1/MmJ
017801 C3Fe.Cg-Fig4plt1/MmJ
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 10sec -
5 - - Go to Step 2-4, 10 times
6 94 15sec -
7 50 15sec -
8 72 10sec -
9 - - Go to Step 6-8, 28 times
10 72 2 min -
11 10 - hold