Strain Protocols

Strain Information
Strain Code: 01XB9
Strain Name: STOCK Brca2tm1Brn/Nci
Gene Symbol: Brca2
Protocol Information
Allele: Brca2<tm1Brn>
Protocol Number: 1
Introductory Comment: The strain Brca2 has loxP sites flanking Exon11. The primer set B012/B013 tests for the presence of the upstream lox site while a second primer set tests for the presence of the downstream lox site located in intron11.

Primers

B012 : 5'-GGc TGT cTT AGA AcT TAG GcT G-3'
B013 : 5'-cTc cAc AcA TAc ATc ATG TGT c-3'

Product Sizes

Primer Combination Product Size
B012 / B013 376 bp    5'Lox
B012 / B013 298 bp    Wildtype

Master Mix

Reagent Final Concentration
10X Buffer 1X
25mM MgCl2 2 mM
2.5mM dNTP Mix 0.2mM
100 µMB012 0.5 µM
100 µMB013 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 0:30
60ºC 0:30 35 Cycles
72ºC 1:00
0ºC
72ºC 3:00
4ºC HOLD

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.