...
1uL mKate, 1uL TREt, 1uL destination vector 1-2
3/8:
Transformed competent E. coli with results of LR reactions
Amplified YFP gene with miRNA target sites, using PCR
3/14:
Gibson assembly -
restriction digest on backbone - 1ug (4.2 uL) DNA (055 plasmid), 1uL Pac1, 2uL buffer, 12.8 uL water; incubate at 37C for 3h
minipreps to extract LR products
3/16:
used nanodrop to measure plasmid concentration in minipreps
ran gel to extract PCR products