2011-05-11 visual protocol

# Thaw competent cells (Lyocomp E. coli) on ice.

# During this time pre-chill sterile centrifuge tubes on ice.

# Gently tap competent cell containers to mix

# Aliquot bacterial suspension in pre-chilled tubes (100 µl per tube).

# To the aliquots add 1 µl of plasmid (RFP PUC19).

# Gently tap centrifuge tubes to mix

# Incubate 30 min on ice.

# During this time pre-heat water bath to 42°C (don’t use heating block).

# Incubate samples at 42°C in the water bath for 60s. Do not shake.

# Immediately chill samples on ice for 2 minutes.

# Add 0.95 ml of room temperature SOC.

# Plate 10 µl of bacterial suspension on antibiotic-containing agar plate. (filter sterilized Kanamycin100 ug/ml?).

# Transfer some bacteria to another antibiotic-containing plate using a loop.

# Incubate plates at 37°C for 18 h.

Done