Strain Protocols

Strain Information
Strain Code: 01XA1
Strain Name: B6.129-Apctm1Rak/Nci
Gene Symbol: Apc
Protocol Information
Allele: Apc<tm1Rak>
Protocol Number: 1
Introductory Comment:

Primers

A001 : 5'-TGc cAG cAc AGA ATA GGc TG-3'
A002 : 5'-TGG AAG GAT TGG AGc TAc GG-3'
A003 : 5'-GTT GTc ATc cAG GTc TGG TG-3'

Product Sizes

Primer Combination Product Size
A001 / A002 400bp    Knockout
A001 / A003 300bp    Wildtype

Master Mix

Reagent Final Concentration
10X Buffer 1X
25mM MgCl2 2.0 mM
2.5mM dNTP Mix 0.2 mM
100 µMA001 0.33 µM
100 µMA002 0.33 µM
100 µMA003 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 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.