Strain Protocols

Strain Information
Strain Code: 01XC9
Strain Name: STOCK Brca1tm2.1Cxd/Nci
Gene Symbol: Brca1
Protocol Information
Allele: Brca1<tm2.1Cxd>
Protocol Number: 1
Introductory Comment: The Brca1 strain is suppose to be heterozygous for deletion of Exon 11. The primer set B004/B006 will give a 600bp band if Exon 11 has been removed while primer set B004/B005 gives a 450bp band and indicates that Exon 11 is present.

Primers

B004 : 5'-cTG GGT AGT TTG TAA GcA TGc-3'
B005 : 5'-cAA TAA AcT GcT GGT cTc AGG-3'
B006 : 5'-cTG cGA GcA GTc TTc AGA AAG-3'

Product Sizes

Primer Combination Product Size
B004 / B005 450 bp    Wildtype
B004 / B006 600 bp    Deletion

Master Mix

Reagent Final Concentration
10X Buffer 1X
25mM MgCl2 2.5 mM
2.5mM dNTP Mix 0.2 mM
100 µMB004 0.33 µM
100 µMB005 0.33 µM
100 µMB006 0.33 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
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.