Media

LB medium

1. weight components

• 5 g/L NaCl
• 10 g/L tryptone
• 5 g/L yeast extract
• (15 g/L agar for plates)

1. fill up to 1 L with deionized water
2. mix well by shaking
3. autoclave
   1. autoclaving tape, caps slightly unscrewed
   2. base of the pot has to be covered with deionized water
   3. close lid
   4. heat level 3 until the pressure valve opens
   5. reduce heat level to 1.5
   6. set timer to 20 minutes
   7. turn heater off
   8. wait until the pressure valve retracts (30-45 minutes)
   9. open, close caps & shake
4. for plates, wait until you can touch the bottle (<60 °C, clean bench!)
5. add antibiotics and shake (gloves!)

Hartmans minimal medium (HM)

Agar Chips

Transformation

Heat Shock

1. thaw cells on ice
2. add 1 µl of plasmid DNA
3. incubate on ice for 30'
4. heat shock at 42 °C for 60
5. incubate on ice for 5'
6. add 200 µl of SOC media
7. incubate at 37 °C for 2 h
8. plate 20  and 200 µl on plates with the appropiate antibiotic

Electroporation

PCR

Colony PCR

QuikChange

Clonings

Restriction Digest

Ligation

Gibson Assembly

SDS-PAGE

For some SDS-PAGEs we used BioRad ready made gels.

The recipe of the self-made SDS is as follows:

3.5x Buffer

Gels