Imperial College/20 August 2008

= 20 August 2008 =

Cloning

 * GFP-3-mutB - Terminator and CAT overnight cultures miniprepped (all 3ml).
 * Single digests (EcoRI) of all registry sequences performed and assayed by gel to check vector size

All major bands correspond to the approximate size of vector and insert. In several of the minis a second band is however visible is most likely to be denatured DNA as a byproduct of doing all the minipreps in one go (and so taking too long)

B.subtilis

 * The transformation protocol 2, | Click link here for protocol was carried out on competent cells that had been grown to an O.D.600 of 1.8.
 * The following concentrations of pDR110 DNA was used from the stock of 40ng/ul; 0, 40ng, 200ng, 400ng. One problem to note is that although the voltage and resistance can be controlled the pulse length cannot. The pulse length controls the length of time that the bacterium is exposed to the voltage. One problem is that if the pulse length drops below about 4msec, the efficiency of transformation is very low. In this set of transformations a range of 3msec to 10msec were seen.

Dry Lab

 * About to complete Tutorial 2
 * Preliminary work on validating the tracking algorithm written by EPFL using video generated from synthetic data
 * Preliminary work on growth curve of Bacillus Subtilis
 * We are aiming to produce a chassis characterisation.