Team:Edinburgh/Notebook/FluidDynamics

Fluid Dynamics

Week 1
Take CBDs out of the registry by transforming, culturing, and then mini prep.


Week 6
08/07
Digest CBDs, MorA and RFP for both N and C terminal fusions.
CBDs: 5.18A, 5.16O, BBa_K863111 and BBa_K863101. For N terminal fusions use AgeI/SpeI and for C terminal fusions use XbaI/NgoMIV. Treat with Antarctic phosphatase.
For MorA and RFP N terminal fusions use NgoMIV abd SpeI, for C terminal fusions use XbaI and AgeI.

PCR purify CBDs. The digest did not really work for the inserts so digest again over night.


09/07
Gel purify the overnight digest.
Ligate each of the 4 CBDs to RFP for both N and C terminal fusions with controls of the CBDs without inserts.


10/07
Transform the ligations.
Plate bacteria for bacterial cellulose in liquid, shaking liquid, and solid at 22ºC.


11/07
Some of the ligations appeared to have worked.


12/07
Inoculate transformants that appeared to have successfully ligated.


Week 7
13/07
Make glycerol stocks of cultures.

Miniprep and diagnostic digest


Week 11

11/07
Measured the chads (chad measurement protocol)
CBD fused with GFP association experiment (association experiment protocol)


13/07
RFP control association experiment
RFP dissociation experiment (dissociation experiment protocol), leave chads in RFP overnight in 4 °C


14/08
Dissociation experiment using chads incubated with RFP overnight
Dissociation experiment using chads saturating in RFP for 5 minutes


17/08
Dissociation experiment of GFP, GFP + 5.21M, RFP


18/08
Dissociation experiment of GFP 5.20M using chads from Whatmann 54, unprocessed bacterial cellulose and processed bacterial cellulose


19/08
ASK MICHELLE


20/08
ASK MICHELLE


21/08
Dissociation experiments of lac I 5.160 RFP C, lac I BBa_K863111 RFP N, lac I BBa_K863111 RFP C


22/08