E coli colony PCR

This is a protocol to do PCR reactions on E. coli cells, directly after a transformation. (See here for colony PCR for yeast cells)
Prepare mastermix, divide 20μl/sample over PCR reaction tubes/wells and add some fresh cells to the mix using pipette tips or toothpicks.
If you actually want to use these cells later, don't forget to maintain the cells on a fresh 'master plate' or in a 96 well plate.

Make sure you add only a few cells. Adding too much might reduce the efficiency of the reaction, resulting in no product. If you're reaction fails, a solution might be to first resuspend your cells in 10μl DNAse free water and then add 1μl of this mix to the reaction.

Do sequencing reaction:
H2O	15.4 μl
Taq PCR Buffer 10X	2 μl
dNTPs (10 mM each)	0.8 μl
MgCl2 (50mM)	0.8 μl
Primer Fwd (10 μM)	0.4 μl
Primer Rev (10 μM)	0.4 μl
Taq polymerase (5U/μl)	0.2 μl

Run using program (bart -> colony PCR)

Number of cycles	Temperature	Duration
1 X	94°C	2 min
35X	94°C	30 sec
	52°C	30 sec
	72°C	1 min 30 sec
1X	72°C	7 min
	7°C	Inf