Team:Groningen/Labwork/31 July 2013

Mirjam

Examined the plates of eYFP and GFP grown on Lb agar + cm plates. This reveals that there are cells growing and the negative control did not show any colonies. Six colonies per type are inoculated in LB medium with cm to perform a mini prep later on.
Examined the plates of the ligation product BBa_k823823+LacI with eYFP and GFP. These plates did not show any colonies, although the negative control did, which is very strange.
The plated B.subtilis strain did grow. So three single colonies are inoculated to perform a transformation reaction.
Made a restriction digestion for RFP and eYFP (EcoRI and PstI).

Chaline & Mirjam

Made a restriction digestion for Pdes (BamHI) and BBa_k823823 + LacI (EcoRI and PstI).
Did a mini prep to obtain more BBa_k823823 + LacI. Made a restriction digestion of these samples to verify the plasmid is correct. Did a nanodrop to measure the concentration of the samples. The samples are fine, with concentrations between 30 and 60 ng/µl.
Run a 0.8% agarose gel on the samples of the restriction of the backbone to do gel purification. Gel purified with a very low concentration of the backbone. Though it is decided to continu with this sample.
Did a

Sander

Made a placmid purification of Munchen backbone with Lacl. Got a concentration of 44,4 and 47,3 ng/ul. (placed in temp box labeled as Mun+Lacl)
Made a spread of the Des knockout on LB agare with Cm 5 ug/ml and Kn 5ug /ml. Left to grow in 37°C.
Made a spread of the CheY knockout on LB agare with Cm 5 ug/ml. Left to grow in 37°C.