Strain Protocols

Strain Information
Strain Code: 01XG9
Strain Name: SWR.129P2(Cg)-Brca2tm1Kamc/Nci
Gene Symbol: Brca2
Protocol Information
Allele: Brca2<tm1Kamc>
Protocol Number: 1
Introductory Comment: The primer set for genotyping Brca2 comes from the paper Cancer Research 62:990-994,2002. The wildtype primer pair and the knockout primer pair both amplify a 700bp product.

Primers

B016 : 5'-GGA GGA GGA GGA GTT GTT GA-3'
B017 : 5'-ATc TcG TTc TcT ccA cTc cA-3'

Product Sizes

Primer Combination Product Size
B016 / B017 700bp    Wildtype

Master Mix

Reagent Final Concentration
10X Buffer 1X
25mM MgCl2 2.0 mM
2.5mM dNTP Mix 0.2 mM
DMSO 1%
100 µMB016 0.5 µM
100 µMB017 0.5 µM
5U/┬Ál Taq Pol 0.025 U/µl
Use 50 - 100 ng of DNA per reaction

Cycling Conditions

(Time in minutes:seconds)
Temperature Time Label
94ºC 3:00
0ºC
94ºC 1:00
60ºC 2:00 35 Cycles
72ºC 1:00
72ºC 3:00

Disclaimer: PCR conditions were optimized using QIAGEN-purified mouse tail DNA and a PTC-200 thermocycler (MJ Research). Optimal conditions may vary with DNA quality or equipment used.