Ligation Reaction - (Apr/26/2008 )

Hi there,

In out lab report it says we should write about "how would we set up a liagation reaction"...is that pretty much just asking for the method?...We basically had solutions containing ligase that that w.o ligase and viewed on agarose gel if the ligation had been successful...so yeah are they pretty much just asking for the method or sumthing else?

b_06er

I think so.

I would write that the ligation reaction contains the vector, the insert, ligase buffer contains ATP and the ligase enzyme and water (if needed). Then you incubate. The incubation can be any degrees between 16 to 37 for 2h-ON. I am not sure you could see the ligation product on the gel (have never tried but it depents how much DNA you put into the reaction). Negative control would be a reaction without the insert. You can try different ratios of vector:insert etc. That`s what I understand??