Strain Protocols

Strain Information
Strain Code: 01XA4
Strain Name: B6.129-Msh6tm1Rak/Nci
Gene Symbol: Msh6
Protocol Information
Allele: Msh6<tm1Rak>
Protocol Number: 1
Introductory Comment:

Primers

M010 : 5'-TGG AAG GAT TGG AGc TAc GG-3'
M011 : 5'-TTA ccT ccT ccA cTG AcG TG-3'
M012 : 5'-cAA Gcc ccT TTc TTT GTT TG-3'
M013 : 5'-Acc AcT Tcc TcA Tcc cTG G-3'

Product Sizes

Primer Combination Product Size
M010 / M011 400 bp    Knockout
M012 / M012 550 bp    Wildtype

Master Mix

Reagent Final Concentration
10X Buffer 1X
25mM MgCl2 2.0 mM
2.5mM dNTP Mix 0.2 mM
100 µMM010 0.25 µM
100 µMM011 0.25 µM
100 µMM012 0.25 µM
100 µMM013 0.25 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
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.