Strain Protocols

Strain Information
Strain Code: 01X66
Strain Name: B6.Cg-Tg(CD207-cre)1Dhka/Nci
Gene Symbol: iCre
Protocol Information
Allele: Tg(CD207-cre)1Dhka
Protocol Number: 1
Introductory Comment: This protocol detects the presence of transgene (Langerin-Cre) using primers y072 and y073. To ensure that the PCR is working properly, an internal control is included using primers y074 and y075. The protocol does not distinguish between heterozygotes and homozygotes.

Primers

y072 : 5'-gag gca aat gat tgg cat tct ac-3'
y074 : 5'-caa atg ttg ctt gtc tgg tg-3'
y073 : 5'-ctg gaa aat tca aga aga gcc t-3'
y075 : 5'-gtc agt cga gtg cac agt tt-3'

Product Sizes

Primer Combination Product Size
y072 / y073 272 bp    transgene
y074 / y075 200 bp    internal control

Master Mix

Reagent Final Concentration
10X Buffer 1X
25mM MgCl2 2 mM
2.5mM dNTP Mix 0.2mM
100 µMy072 1.0 µM
100 µMy074 1.0 µM
100 µMy073 1.0 µM
100 µMy075 1.0 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 0:30
55ºC 0:30 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.