I'm currently looking for a protocol to cut a plasmid and ligate the inserts into another.
(I asked a proff a while ago if I could get SalI. Now he replied 'yes'. So I'm cutting out the light gene casette of pVIB ( http://images.carolina.com/images/en_US//local/products/detail/211447_bit.jpg - only one cutting site?) and insert it into a kanamycin resistant plasmid.)
I won't do electrophoresis. Just plate it on Kanamycin, then some collonies will glow, while others will not. Then I take a glowing one, grow it, and do a miniprep.
But how much RE will I need, how long to wait until cut, how much plasmid and how much DNA ligase?
Would be terrific if somebody could send me a protocoll!!
Thanks!!
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