I am performing a sequential restriction for a plasmid with two enzymes that cut once.
I am using two enzymes from different companies and what i have done after restriction, i just heat inactivate the enzyme and used an amount of the first reaction to the second reaction where i added the buffer that matches the second enzyme.
Is it necessary to purify the first reaction or it works with heat inactivation since i thought about what happens to the buffer from the first reaction? would it affect the second reaction even if i heat inactivated the enzyme
Edited by thegene, 01 March 2010 - 03:04 AM.