Team:UNIK Copenhagen/Signe Notebook
From 2013.igem.org
Notebook
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Week 1 - July 8th-14th
Culturing of magnetotactic bacteria (MTBs)
Making base media for growing magnetotactic bacteria. This media was also used to make agar plates.
MS-1 inoculated in liquid base medium. Falcon tubes were flushed with the nitrogen before use and incubated at 28 degrees.
Plates were kept in a closed container with an Oxoid AnaeroGen pad.
Assembly of constructs
Glycerol stock of E. coli with pBBR1MCS-2 was thawed to grow on these plates.
Primers for MamC (MS-1 strain) was ordered.
Week 2
Assembly of constructs
Overnight liquid cultures of pBBR1MCS-2 and eGFP plasmid (Adam Takos)
Touchdown PCR amplification of MamC from MS-1 gDNA (both Phusion and Hotmaster) and of eGFP (only Hotmaster).
Gel purification of bands from PCR amplification and subsequent nanodrop. Due to poor yield from this extraction gradient PCR was performed (56-68 degrees).
Traditional cloning of eGFP fragments into pDRIVE.
Transformation of eFbFP-pEX-A (synthesized gene, Eurofins) and eGFP-pDRIVE.
Gel purification of bands from PCR amplification and subsequent nanodrop. Due to poor yield from this extraction gradient PCR was performed (56-68 degrees).
Picture 12.20
Traditional cloning of eGFP fragments into pDRIVE.
Transformation of eFbFP-pEX-A (synthesized gene, Eurofins) and eGFP-pDRIVE.
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