Team:Freiburg/Notebook/6 September

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Commons

Miniprep

Investigators: Sandra

Yesterday LB was inoculated with strains (S14, S39-S44 etc.). Miniprep of strains.

Digestion: 2A-assembly

Investigators: Sandra

Digestion of:

• S14 (GFP)

with XbaI and PstI

• S39 (PR1)
• S40 (PR2)
• S41 (PR3)
• S42 (PR4)
• S43 (PR5)
• S44 (PR6)

with SpeI and PstI

incubation for 6 hours at 37°C.

green light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME

blue light receptor

Trafo

Investigators: Sandra

Trafo of ligated parts of the 2A-assembly.

Sequence analysis

sequence analysis of our lovtap-Notgate was neagtive:-( So we´ll try the gibson assembly.

PCR

Investigators: Sophie PCR

PCR-Mixture for one Reaction:

For a 50 µl reaction use

Digestion with Dpn I and purification with PCR purification kit What program do you use?

First 20 cycles touchdown 69°C -0.4/cycle

next 10 cycles touchdown 72°C -0.2 per cycle

To confirm the PCR-Product has the correct size, load 2 µl of the sample onto an agarose-gel.

How did you label the PCR-Product, where is it stored and what do you do next?

red light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME

Lysis cassette

Troubleshooting of the modified Lysis genes K124017

Investigators:Theo

Minipreps of the first inoculations were applied on a 1% agarose gel without digesting them to see if they have an insert of the modified Lysis genes K124017. Nr1, 5 and 7 were sent for sequencing since the size seems to be about right.

• Note: S4 is the B0034 RBS (ribosome binding site), S15 is the non-modified K124017 Lysis Cassette without RBS

Precipitator

NAME OF YOUR EXPERIMENT

Investigators: NAME