50 ng of each piece of DNA being joined (use nanodrop to find concentration in ng/ul, then divide 50 by that concentration to find the required volume of DNA)
100ng Donor
2ul 10X T4 Ligase Buffer
2ul 10X BSA
1ul BsaI-HF
1ul T4 Ligase HC
fill to 20uL with SDIH20
-------------
20ul total
protocol EBGG
37 5min
50X:
37 2.5min
4C 0.5min
16C 5.5min
37C 10 min
80C 20 min
4C hold
takes 8+ hours
(check protocol by looking up the paper or other online GG protocols)