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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)