FVB/N-Tg(Fabp1-Cre)1Jig PCR Protocol |
| Strain: FVB/N-Tg(Fabp1-Cre)1Jig |
Strain Code: 01XD8 |
| Protocol Number: 1 |
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| Introductory Comment: |
This strain contains a Cre recombinase gene linked to the rat fatty acid binding protein 1 (Fabp1) promoter. The genotying strategy is to detect this transgene with primer F006, which is specific for the Fabp1 element, and C031, which is specific for the Cre recombinase gene. |
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Primers:
F006 : 5'-tcg ttg acc att gct ctc ag -3'
C031 : 5'-cat cac tcg ttg cat cga cc -3'
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Product
Sizes:
| Primer
Combination |
Product Size |
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| F006 /C031
|
214 bp Fabp1-Cre Tg |
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Master Mix
| Reagent |
Final
Concentration |
| 10X Buffer |
1X
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| 25mM MgCl2 |
2.0 mM
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| 2.5mM dNTP Mix |
0.2 mM
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| 100 µM F006 |
0.5 µM
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| 100 µM C031 |
0.5 µM
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| 5U/µl Taq Pol |
0.015 U
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 |
| Use 50 - 100 ng of DNA per reaction |
Cycling Conditions (Time in minutes:seconds)
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Temperature
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Time
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Label |
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94ºC
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3:00
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94ºC
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0:30
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60ºC
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0:30
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35 Cycles |
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72ºC
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0:30
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72ºC
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3:00
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4ºC
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HOLD
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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.
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