Strain Protocols

Strain Information
Strain Code: 01XF2
Strain Name: 129-Nf1tm1Fcr/Nci
Gene Symbol: Nf1
Protocol Information
Allele: Nf1<tm1Fcr>
Protocol Number: 1
Introductory Comment:

Primers

N010 : 5'-GTA TTG AAT TGA AGc Acc TTT GTT TGG-3'
N012 : 5'-cTG ccc AAG GcT ccc ccA G-3'
N011 : 5'-GcG TGT TcG AAT TcG ccA ATG-3'

Product Sizes

Primer Combination Product Size
N010 / N012 200 bp    Wildtype
N011 / N012 350 bp    Knockout

Master Mix

Reagent Final Concentration
10X Buffer 1X
25mM MgCl2 2.0 mM
2.5mM dNTP Mix 0.2 mM
100 µMN010 0.33 µM
100 µMN012 0.33 µM
100 µMN011 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
94ºC 1:00
65ºC 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.