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

• mix well by shaking

1. 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
2. for plates, wait until you can touch the bottle (<60 °C, clean bench!)
3. add antibiotics and shake (gloves!)

Hartmans minimal medium (HM)

Agar Chips

Figure 1 This is our team logo

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