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Team:UGA-Georgia/NotebookMethanococcus
From 2013.igem.org
Methanococcus Lab
Instructor: ZHE LYU
February 2013
-Training on anaerobic skills, i.e. use of anaerobic glassware, gassing chamber, anaerobic chamber, etc.
March 2013
-Purification of GS, AT and GS+AT via revival of frozen stocks and plating of sub-cultures.
April 2013
-Further continuation of purification of GS, AT and GS+AT via picking colonies, creating sub-cultures and plating.
-Test of Puromycin strength.
-Creation of frozen stocks of purified GS, AT and GS+AT transformants.
June 2013
-Extraction of Geraniol from extra-cellular and intra-cellular content of samples and preparation for GC/MS evaluation
-Sequencing of pAW50-mCherry vector
-Analysis of pAW50-mCherry sequence
-Transformation of pAW50-mCherry vector into Methanococcus
-Testing Fluorescence of pAW50-mCherry
-Purification of pAW50-mCherry cultures via plating of transformants, and creating sub-cultures of colonies picked
-Creation of frozen stocks of pAW50-mCherry in Methanococcus
July 2013
-Creation and analysis of the "killing & inhibiting" experiment where we test the maximum amount of product a 5ml culture of Methanococcus can tolerate in cultures with high OD (killing of grown cells) and low OD (inhibiting of growth).
-Expanded upon the original extraction protocol for higher efficiency of extraction of geraniol from Methanococcus cultures.
- GC/MS Evaluation and analysis (see results tab)
-Innovation of an adapter to allow the use of syringe needles on micropipettes.
August 2013
-Revival and PCR of all GS and AT frozen stocks to confirm insert.
--Extraction of Geraniol from extra-cellular and intra-cellular content of all GS frozen stocks and preparation for GC/MS evaluation
September 2013
-Transformation of BBa-k1138002 into Methanococcus
-Testing Fluorescence of BBa-k1138002
-Purification of BBa-k1138002 cultures via plating of transformants, and creating sub-cultures of colonies picked
-Creation of frozen stocks of BBa-k1138002 in Methanococcus
-GC/MS Evaluation and analysis (see results tab)
