Table S2.

Timetable for generation of biGBac expression constructs

DayExperimental stepsTime*
Day 1PCR GECs∼8–10 h
PCR “cleanup,” determine DNA concentration, agarose gel electrophoresis
Step 1 Gibson assembly reaction
Transformation of E. coli
Day 2Inoculate E. coli overnight cultures∼20 min
Day 3“Miniprep” DNA isolation∼6–8 h
SwaI/PmeI digests, agarose gel electrophoresis
Submit for DNA Sanger sequencing
Day 4Analyze sequencing results∼6 h
Step 2 PmeI digest and Gibson assembly reaction
Transformation of E. coli
Day 5Inoculate E. coli overnight cultures∼10 min
Day 6Miniprep DNA isolation∼4 h
SwaI/PacI digests, agarose gel electrophoresis
  • * Times are estimated for the generation of five pBIG1 and one pBIG2 construct. The generation of multiple constructs in parallel can require longer periods of time.