From 2014.igem.org
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Notebook |
February
Week 1
Introduction to anaerobic facilities for new members
Week 3
Making of NaS solution.
Making of formate media.
Week 4
Creation of transformation buffer (TB).
First geraniol extraction efficiency test with balch tubes.
March
Week 1
Creation of geraniol standards for gas chromatography (GC) analysis/calibration through serial dilutions.
Week 3
Creation of more formate media.
Creation of general salts solution.
Creation of glycylglycine buffer.
April
Week 1
Learning about ribosome binding site library (RBS) and our metabolic Optflux model.
Week 2
Made agar plates
Creation of fresh wild type (WT) stocks.
Week 3
Created diluted puromycin
May
Week 4
Inoculation and revival of frozen stocks.
June
Week 1
Determined the optical density (OD) of the previously revived cultures.
Week 2
Restriction digestion, purification, and ligation of pMEV4 and PCR products (C1 and C2).
Transformation by heat shock, plated the cells, made an agarose gel and ran the products on it.
Picked colonies from the previous day of plating.
Plasmid extraction, digestion, gel electrophoresis, and verification.
Week 3
Revival of pAW50-mCherry frozen stock.
Revival of S0001 (WT) frozen stock.
Week 4
Prepared the revived cultures for fluorescence microscopy.
Fluorescence microscopy training.
Anaerobic transformation of RBS libraries 1-12 into M. maripaludis.
Puromycin enrichment of transformants.
Made glycerol stocks of original transformants.
Fluorescent microscopy of transformants/pAW50-mCherry/S0001.
July
Week 1
Took ODs of the native RBS and library 1 and 2 (12; L-1; L-2 respectively).
Plated the 12, L-1, and L-2 cultures.
Dispensed media to test tubes.
Picked colonies form the plates.
Week 2
ODs from last weeks picked colonies were taken.
Made frozen stocks of RBS colonies,
Sub-cultured parent strains.
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