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Tempel Lab Protocols

BDNF GENOTYPE

You will Need

Mouse Tail Digests

2 Primers:
IMR 1846
IMR 1847

PCR Reagents:
Sterile water
10x buffer w/ MgCl2
10mM Nucleotide mix
FastStart Taq

Primer Sequences

IMR 1846 TGTGATTGTGTTTCTGGTGAC
IMR 1847 GCCTTCATGCAACCGAAGTATG

Procedure

Single reaction.

PCR Rxn

Sterile Water

17.8 ul

10x PCR buffer w/MgCl2

2.5ul

10mM Nucleotide mix

0.5ul

IMR 1846 20 uM

1ul

IMR 1847 20 uM

1ul

FastStart Taq

0.2ul

Tail Lysate  (1:10 diluted)

2ul

Total Volume

25ul
   
   
   
 

PCR Protocol

94 deg C

4:30

  

94 deg C

0:30

  

65 deg C

1:00

x35

72 deg C

1:00

  

72 deg C

7:00

  

4 deg C

Hold

  
     

Gel
2 % Agarose + 2 ul EtBr (10mg/ml)

Bands
Wildtype: 437 bp
Flox1: both
Fox2: 487 bp