Primers:

lzr forward (bwd385):  act cgg cgg act ctc gca agc

lzr reverse (bwd386):  ggc tct cgt cgg tga tgg cca tga tct tct

product size:  128bp

XbaI fragment sizes:  30bp and 98bp

Method:

1) Fin clip and add 50ul Lysis Buffer and 5ul 10mg/ml  Proteinase k per embryo.

2) PCR reaction:

11.8ul  ddH2O

2ul   10X PCR Buffer

1ul   2.5mM MgCl2

1ul   5mM dNTPs

1ul   5um forward primer

1ul   5um reverse primer

2ul   DNA

0.2ul  Taq Polymerase

PCR Profile:

94C  1 min

94C  20 sec

65C  20 sec

72C  20 sec

Goto step 2 40X

4C soak

3) Restriction Digest:

10ul PCR Product

2ul 10X Buffer

0.5ul XbaI

7.5ul  ddH2O

incubate at 37C for 3hrs

4)  Separate fragments using a 2% Agarose gel