PCR Reaction for GATEWAY Cloning

The template, if it is a plasmid, should be linearized to reduce backgrownd colonies that contain the original vector in the transformations of the BP and LR reactions

	Init. Conc	Fin. Conc	Fin.Vol
DDW	_	_	34
Buff.	x10	x1	5
Buff Enh.	x10	x1	5
dNTP's	10	0.2	1
Mg+2	50	1	1
For. Primer	10	0.25	1.25
Back. Primer	10	0.25	1.25
Taq Pol.	_	_	0.5
Template	_	_	1
Tot. Vol.	_	_	50

 

Step	1	2	3	4	5	6
Conditions	2' 94oc	15'' 94oc	1' 55oc	1' 68oc	10' 68oc	4oc

Repeat steps 2-4 30 cycles.