Difference between revisions of "Team:TUDelft/Notebook"

Revision as of 12:05, 27 September 2019

Sci-Phi 29

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Cloning

Testing of Plasmid

April

May

June

July

August

Goal:	Assembly all the different parts with MoClo
Protocols Used:	Nanodrop DNA quantification, Plasmid Isolation, MoClo, E.coli Heat Shock Transformation, KOD Extreme PCR, Colony Polymerase PCR (GoTaq), E. coli Glycerol Stock, DNA Gel Electrophoresis Sequencing
Plan:	For Orthogonal Replication, first, all the level 0's were constructed and then assembled in level 1 to make the following constructs: 0.1 T7 p - U RBS - DNAP - WT 0.5 T7 p - U RBS - DNAP - WT WT T7 p - U RBS - DNAP - WT 0.1 T7 p - U RBS - TP - WT 0.5 T7 p - U RBS - TP - WT WT T7 p - U RBS - TP - WT 0.1 T7 p - U RBS - p5 - WT 0.5 T7 p - U RBS - p5 - WT WT T7 p - U RBS - p5 - WT 0.1 T7 p - U RBS - p6 - WT 0.5 T7 p - U RBS - p6 - WT WT T7 p - U RBS - p6 - WT 0.1 T7 p - U RBS - GFP - WT 0.5 T7 p - U RBS - GFP - WT WT T7 p - U RBS - GFP - WT
Results:	All composite parts are confirmed by sequencing.

September

Goal:	Add characterization for the parts page and see if the different promoters strength are visible
Protocols Used:	PURE System, Protein Gel, Mass Spectrometry,
Plan:	The following constructs were added to the PURE system and then run on a protein gel: 0.1 T7 p - U RBS - DNAP - WT 0.5 T7 p - U RBS - DNAP - WT WT T7 p - U RBS - DNAP - WT 0.1 T7 p - U RBS - TP - WT 0.5 T7 p - U RBS - TP - WT WT T7 p - U RBS - TP - WT 0.1 T7 p - U RBS - p5 - WT 0.5 T7 p - U RBS - p5 - WT WT T7 p - U RBS - p5 - WT 0.1 T7 p - U RBS - p6 - WT 0.5 T7 p - U RBS - p6 - WT WT T7 p - U RBS - p6 - WT After this, a remainder of the sample was analyzed with Mass Spectrometry.
Results:	Add link

October

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