Team:Groningen/Labwork/7 August 2013

Mirjam

A check on the plates with CFP revealed that no colonies are present.
A check on the plates for ΔCheY revealed colonies. The ΔDes plates did not reveal any colonies.
Check of the restriction digest made 06-08 of CheY, and the two tubes of CFP. This reveals that CheY and one of the CFP tubes is totally absent.
Made a restriction digestion of GFP/YFP with Xba and PstI and did gel purification.
Made a restriction digestion of CFP with EcoRI and PstI and did gel purification.
Did a gel purification on BBa_k823823 restricted with EcoRI and SpeI.
Made a restriction digestion of CheY with BamHI and SpeI. Heat inactivated the reaction.
Made a ligation between Pdes and CheY and heat inactivated the reaction.
Made a ligation between Pdes/CheY and the backbone and Pdes/CheY/eYFP and the backbone.
Made a ligation between CFP and BBa_k823823 + LacI.
Did a transformation to E. coli of all of the ligation reactions. And plated the reaction of CFP also on IPTG plates.

Chaline

Measured the concentration of three mini prep tubes of CFP and one of the restriction digests.
Measured the concentration of the restricted GFP, YFP and BBa_k823823.
Run a gel with the (purified) restrictions of BBa_k823823, CFP, GFP, YFP and CheY. Only the one with GFP does not show any bands.

Claudio

The Bacillus Subtilis biofilm grew on 2x SG medium after 4 and 5 days of incubation respectively.
2x SGG is prepared in order to grow biofilm between liquid-air interface.

Sebas

Did colony pcr on delta des and cheY transformants (made by Mirjam)-> no bands visible on gel indicating that they
did not contain the amplicon.

Transformed munichbackbone+hyperspank promoter+ GFP/YFP (linearized with scaI) to B. subilis 168