Stock Number:012871
Strain Name:STOCK Pik3r1tm1Lca/J
Pik3r1tm1Lca
Pik3r1tm1Lca
Standard PCR
1.0
07-July -2010 25-August -2010
Expected Results:

Mutant = 338 bp
Heterozygote = 148 bp and 338 bp
Wild type = 148 bp
Gel Image 
Image Image file
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Protocol Primers 
Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type
10744 - GGT TTC TTA CTT TAG ACG GAG CTG - Forward
10745 - CAG ACA GAC CAA CCT AGA GAT TAG G - Reverse
Reaction/Components 
Reaction Component Volume (µl) Final
Concentration		 Total Volume (µl)
ddH2O 4.92 - 4.92
5 mM DNA Loading Dye 1.66 0.69 mM 1.66
DNA 2.00 0.00 2.00
2.5 mM dNTP 0.96 0.20 mM 0.96
10 X KAPA Rxn Buffer 1.20 1.00 X 1.20
20 uM 10744 0.60 1.00 uM 0.60
20 uM 10745 0.60 1.00 uM 0.60
5 U/ul KAPATaq DNA Polymerase 0.06 0.03 U/ul 0.06
Cycling 
Step # Temp °C Time Note
1 94 3 min -
2 94 30 sec -
3 66 30 sec -
4 72 30 sec -
5 - - Go to step 2-4, 35 times
6 72 2 min -
7 10 - hold
This genotyping protocol is used for the following strains:
 
Stock Number Strain Name
012871 STOCK Pik3r1tm1Lca/J
Genotyping resources and troubleshooting
This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.