Team:Freiburg/Notebook/3 September

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green light receptor

Testdigestion of 2a-assembly

Investigators: Julia
Continuing from 31 of Aug. [1]
Before Miniprep of 20 different colonies.
Each minipreped DNA-sample (3μl)was digested with EcoRI and PstI.

Result:

the samples

46b= ho1 with a RBS (BBa_B0034)

50b= RBS(BBa_B0034)+pcyA+terminator

64b= Promotor (BBa_J23110)+CcaS

67b= romotor (BBa_J23110)+CcaR

were send to GATC for sequencing

3A-assembly:Digestion

Investigators: Sandra

Digestion of:

InsertI:

• 64
• 46

with EcorI and SpeI

InsertII:

• 67
• 50

with XbaI and PstI

Incubation for 6 hours at 37°C. Vector pSB1K3 was already digested, it only needs to be digested with antarctica for one hour.

3A-assembly:Ligation

Investigators: Julia

blue light receptor

Stop of ligation

Investigators: Sandra

Inactivation of ligase at 80°C for 20 minutes.
Stored in the blue light box, labelled: ♥C3 Lig., and Not C3 Lig.

To do: Trafo

Transformation

with ♥C3 Lig. and Not C3 Lig

Investigator: Sophie

red light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME

Lysis cassette

NAME OF YOUR EXPERIMENT

Investigators:NAME

Precipitator

Picking clones

Investigators: Sandra

Only few colonies grew on two plates. These clones were picked and incubated in LB with amp at 37°C.