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 3
Culturing of MTBs
making charcoal media and plates. MS-1 and MSR-1 was inoculated. Liquid cultures were setup in Hungate tubes 200ul, 400ul and 600ul of each strain (MS-1 and MSR-1). Charcoal plates were inoculated as well with both strains.
Assembly of constructs
Colony PCR of eFbFP-pEX-A and eGFP-pDRIVE. Only eFbFP was succesful.
Overnight cultures were made of colony 1-4 (eFbFP-pEX-A).
Miniprep of eFbFP-pEX-A cultures and nanodrop. Samples sent for sequencing.
Colony PCR of eGFP-pDRIVE was repeated. Still negative.
Cloning and transformation was also repeated. CloneJET PCR cloning kit was used to create eGFP-pJET (pJET was chosen instead of pDRIVE). Not successful.
Gradient PCR of eGFP premade vector (56-65 degrees). Gel electrophoresis was carried out and the bands were purified from the gel.
Cloning of eGFP into pDRIVE. Not successful.
Cloning of eFbFP into expression vector pBBR1MCS-2.
Colony PCR of eGFP-pDRIVE and eFbFP-pBBR1MCS-2. Gel electrophoresis only reveal one colony to be successful (eFbFP-pBBR1MCS-2 no. 17).
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