Take DNA, determine concentration and length (using NanoDrop and Geneious/Gel)
Determine ng of DNA needed to create 40 fmol solutions.
ng = amount in fmol x length of plasmid in bp x 0.00066 (ng / fmol )
(Check formula using Google)
100ng DNA
5ul each gBlock @ 40fmol each
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)