Changes

Jump to: navigation, search

Restriction Enzyme Based Cloning

1 byte added, 14:40, 7 May 2009
m
no edit summary
#Digest both vector (~1ug) and insert in appropriate buffer 1-2h at 37C. Add 1 uL of CIAP to the vector for the last ~10min at 37C.
#Gel purify both vector and insert (Qiagen kit).
:*Run out on gel (see [[Preparing an Agarose Gel]])
:*On a gel box cut out appropriate bands and place in clean eppendorf tubes. Try to limit the exposure to UV light
:*Weigh out gel piece. For each mg add 3 uL of QG (orange) buffer. For example, for a 0.1g piece add 300 uL buffer.

Navigation menu