Ligate pBR322, GFP(LVA), adapters

 

Partly from Sambrook, 1.68.

Materials:

Methods:

1. set up tubes to anneal and ligate adapters, GFP(LVA), and pBR322 vector:

   

2. Incubate solution for 5 min to melt any cohesive termini that have reannealed. Chill the mixture on ice to .
3. add the following to tube (for a total of ):

   

4. Incubate the reactions for 4-16 hours (rather than suggested 4-16 hours ).

Results: ( 2/9/99)
Let ligation reaction go for about 18 hours. Then, used 1/2 of each reaction in a gel purification. Left other 1/2 on ice.