PCR Colony Screening

Reagents and Materials
Colonies on a plate
Sterile toothpicks
ddH20
PCR tubes
LB Plate containing appropriate selective antibiotic and a grid
DNA Polymerase
10 PCR Buffer
10mM dNTPs
primer set
thermal cycler
agarose gel

1. Touch toothpick to a colony. Touch toothpick to 35ul ddH20 in a PCR tube.
2. Streak a sectored agar plate with the toothpick.
3. Make a PCR Master mix and add 15ul to each PCR tube inoculted with a colony.

Per reaction:
10X PCR Buffer	5ul
10mM dNTPs	1ul
primer set	30um each
DNA Polymerase	0.5
dH20 to	15ul

1 minute @ 95°C
2 minutes @ 50°C (or primer Tm-5)
2 minutes @ 72°C
35 cycles

4. Run 10 ul of product on an agarose gel alongside appropriate controls.