Parts taken from the registry and transformed by electroporation into XL1-Blue E.coli along with XL1-Blue controls and grown on Kanamycin and Ampicillin plates. Parts taken:
The basic principle of this protocol is that competent cells are prepared by growing a culture to a high O.D.600 and then performing electroporation on these competent cells.
Today we grew the competent cells, we used an O.D.600 of 1.5 before we harvested them. In addition electroporation was carried out using the plasmid pDR110. Previously we had mini-preped this to give a stock of 40ng/ul. We transformed with 40ng, 120ng, 200ng and 400ng in a volume of 10ul (water was used to make up to 10ul).
Transformed cells were plated out and placed into the 30oC incubator.
Dry Lab
Completed A More Complex Example of Bayesian Parameter Estimation of Tutorial 2.
Sourced for better tracking algorithm, found SpotTracker which is more accurate than ParticleTracker.
Went for microscope training, obtained 2 x videos on B.Subtilis motility, stored on server with James.
Discussion with Dr. Suhail ( from Structural Bioinformatics Group,Imperial College London) on the computing requirements to analyse the cells motility. We will be allocated a linux 64-bit workstation, that we will be able to access remotely via SSH.
Microscope
Chris, Clinton, James and Prudence received microscope training on Widefield 1, recording two 60s videos of B.subtilis swimming for analysis
Determined that best results would be obtained by bringing a B.subtilis overnight culture to the microscope facility and diluting 100 fold.