Bio-Rad Ligation and Transformation Module - Manual (Page 32)

Transformation – Quick
Guide
Preparation for Competent Cells
1. Label an LB Amp IPTG agar
plate with your initials and place
at 37°C.
2. If not already done, pipet 1.5 ml
C-growth medium into a 15 ml
culture tube. Label with your
initials and warm to 37°C for at
least 10 min. Also ensure your
starter culture is shaking at 37°C.
3. Pipet 150 µl of fresh starter
culture (inoculated yesterday)
into the pre-warmed C-growth
medium and place in shaking
water bath at 37°C.
4. Label a 1.5 ml microcentrifuge
tube with your initials and
"competent cells."
5. Prepare transformation (TF) buffer by
combining 250 µl of transformation
reagent A and 250 µl of
transformation reagent B into a
tube labeled transformation
buffer. Keep on ice.
6. After a 20–40 min incubation,
transfer the actively growing
culture in the C-growth medium
to your competent cells
microcentrifuge tube.
28
37°C shaking
water bath
Starter
culture
C-growth
medium
Ice B
A
37°C incubator