Stock Number:017476
Strain Name:129S-Ucp1tm1Kz/J
Ucp1tm1Kz
Ucp1tm1Kzalternate1
Separated PCR
1.0
22-March -2012 11-April -2012
Expected Results:

Mutant = ~500 bp
Heterozygote = 211 bp and ~500 bp
Wild type = 211 bp
Gel Image 
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Protocol Primers 
Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type
14353 - GGG GTA GTA TGC AAG AGA GGT G - Common
14354 - CCT TTA TAC CTA ATG GTA CTG GAA GC - Wild type Reverse
14355 - CCT ACC CGC TTC CAT TGC TCA - Mutant Reverse
Reaction/Components 
Reaction Component Volume (µl) Final
Concentration		 Total Volume (µl)
20 uM 14354 0.30 0.25 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 14353 0.30 0.25 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 14353 0.30 0.25 uM 0.30
20 uM 14355 0.30 0.25 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
003124 B6.129-Ucp1tm1Kz/J
017476 129S-Ucp1tm1Kz/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 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