Harvard BioDesign/9 June 2014

From 2014.igem.org

Making Plates!

YESCA + 100ug/mL Carb + 25ug/mL CR + 3ug/mL CBB + 0.5mM IPTG YESCA + 25ug/mL Cm + 25ug/mL CR + 3ug/mL CBB + 0.5mM IPTG

Pure LB LB+ 100ug/mL Carb LB + 15ug/mL Cm

Carbenicillin plates marked with 1 black line on side of plate Chloremphenicol plates marked with 1 blue line on side of plate

Transformations:

iGEM transformation efficiency plasmids were transformed into Turbo cells using heat shock high efficiency transformation protocol 5 samples @ 0.5ng/uL, 5 ng/uL, 10ng/uL, 20ng/uL, 50ng/uL.

We attempted to transform the plasmids below in Turbo cells using electroporation. Since Turbo cells are chemically competent, not electrically competent, they arced (meaning the electroporation failed); however, we continued with the protocol, recovering them with SOC and plating them. We then redid the transformation of these plasmids using the heat shock protocol.

pBbE1a (- ctrl) pBbE1a CsgA + PUC19 (-ctrl)

Plating

Plated pBbE1a (- ctrl), pBbE1a CsgA +, and PUC19 (-ctrl) electroporation and heat shock transformed Turbo cells onto 2 LB+Carb plates each plating 10uL and 200uL with glass beads for a total of 6 plates.

Plated iGEM transformation efficiency plasmid transformed cells each onto 2 LB+Cm plates @ 10uL and 200uL with glass beads for a total of 10 plates.

All plates put in 37C warm room to grow up overnight