Miniprep setup & Gel #2 (inconclusive)

Contributors: Esther, Chloe

Chemically competent cell prep — completed

Inoculated 100 ml LB with 1 ml overnight culture, grown to OD₆₀₀ 0.3–0.6 (~2h 55m). Standard MgCl₂/CaCl₂/glycerol competent cell prep, aliquoted and flash-frozen at −80 °C.

16th June plate results

• Chloramphenicol CxnA-1: 3 colonies
• Chloramphenicol CxnA-2: 1 colony
• Chloramphenicol CxnA-Hist: many colonies
• Ampicillin CxnA-Hist: strange growth
• Kanamycin CxnA-Hist: no colonies

Nanodrop

Miniprep setup

4 colonies selected, grown in 5 ml LB + chloramphenicol overnight for plasmid extraction.

Gel #2 — verify plasmid DNA and cut sites

Run at 100 V for 45 min.

Result: gel results not good — bands did not appear.