2x5 mL LB medium were inoculated with 50 µL DH5α, aliquoted for a preculture and incubated overnight (16,5h) at 37 °C.

250 mL were inoculated with 5 mL of preculture and incubated until they reached an OD of 0,5. After centrifugation, the cell pellet was washed with HTP buffer and resuspended in 3ml HTP buffer. In the end, 225 µL DMSO was added to an end concenctration of 6-7%. The 50 µL aliquots were frozen in N2(l) and stored at -80 °C.

1 µL plasmid with a amp-resistence was put on ice for 10 min. A heat shock was induced for 45 s at 42 °C. The sample was put on ice for another 10 min and then incubated in 200 µL LB for 1h before it was plated onto LB-, ampicilin- and canamycin-plates.

50 mL LB-medium (with ampicilin) were inoculated with E.coli BL21 PlyS and grown overnight.

Plates from 26.03.2014

Inoculation of LB-plates with E.Coli DH5α with PSG1164.

Miniprep according to the miniprep kit protocol (Omega) with 2x6 mL preculture.

DNA-concentration:

• 1. 233,2 ng/µL
• 2. 261,3 ng/µL

Digestion scheme:

Incubate samples for 60 min at 37 °C, then induce a heat shock for 2 min at 45 °C. 5 µL marker and 10 µL of the samples are loaded onto the SDS-gel.