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Team:TorontoMaRSDiscovery/Notebook
From 2009.igem.org
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#*1% gel: 0.5 g agar mixed with 50 ml of 1X TBE | #*1% gel: 0.5 g agar mixed with 50 ml of 1X TBE | ||
#*10X TBE diluted with 5 ml TBE + 45 ml ddH20 – did not use all 50 ml for 1 gel (gel will be too thick – make 2 instead) | #*10X TBE diluted with 5 ml TBE + 45 ml ddH20 – did not use all 50 ml for 1 gel (gel will be too thick – make 2 instead) | ||
| - | + | #*''Loading Dye'': add glycerol solution to 25 mg bromophenol blue then add dH20 to 10 ml (to make 6X) | |
| - | + | #*Running gel: match wells to black side, run at 120 mA | |
| + | #Visualize Gel in UV | ||
| + | ##Turn power on | ||
| + | ##Gel in machine face up | ||
| + | ##Close door securely | ||
| + | ##Turn white light on | ||
| + | ##Adjust zoom, contrast, focus from black dial on top of machine | ||
| + | ##Turn white light off (turns on UV) | ||
| + | ##Press ‘live’ toggle – acq. Should be 0.4 sec. | ||
| + | ##Print if desired or save on floppy disk | ||
| + | ##Turn power off | ||
| + | ##Dispose of gel in proper container | ||
| + | ##Close door | ||
| - | + | =May 25, 2009= | |
| - | + | #Made overnight bacteria culture (200 ml LB: 20 microliters DB3.1) | |
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| - | 2009 | + | =May 26, 2009= |
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Revision as of 15:48, 4 September 2009
Contents |
April 27, 2009
- Received from Rosa (SPiT):
- TM0785
- Plasmid containing encapsulin
- Recommend transfect into bacteria and re-sequence
- See email note regarding sequence error
- 0.5 microliters TMG DNA 100 microgram/microliter
- Use 0.4 microliter for 50 microliter PCR reaction
- This is thumotoga maritime genomic DNA for purpose of re-cloning
- TM0785
- Microcentrifuge tubes 1 and 2 placed in -20 freezer
May 15, 2009
- pH buffers received from VWR Mississauga
- pH 4 buffer (red) 500 ml
- pH 7 buffer (yellow) 500 ml
- pH 10 buffer (blue) 500 ml
Above are used for pH/mV Meter calibration
- pH/mV meter calibrated according to manual – recorded in index
- Ethanol solution (70%) made from 85% ethanol
May 19, 2009
- 2L of TE buffer made (10X TE)
- Recipe for 2L from stock solution (10X TE)
- a) 10 ml 1M Tris-HCl pH 7.5-80.0 x 2 = 20 ml
- b) 2 ml 0.5 M EDTA pH 8.0 x 2 = 4 ml
- c) 988 ml ddH20 x 2 = 1976 ml
- To make Tris-HCl, Tris base is used and adjusted to desired pH using HCl
- 1 M Tris base is required of which 20 ml is required – thereby 2.4228 g Tris base (due to scale limitations, 2.42g of Tris Base was used
- Recipe for 2L from stock solution (10X TE)
- 500 ml of 1 M Tris Base made
- mass of Tris used = 1M/1000 ml x 500 ml x 121.14 g/mol = 60.57 g
- volume of water used = 500 ml
- 250 ml of 0.5 M EDTA solution was made
- mass of EDTA used = 36.53 g
- observations: EDTA did not dissolve in ddH2O on heat and being stirred
May 21, 2009
- Retrieved autoclaved ddH20, glycerol solution
- Gel Electrophoresis (test run)
- 1% gel: 0.5 g agar mixed with 50 ml of 1X TBE
- 10X TBE diluted with 5 ml TBE + 45 ml ddH20 – did not use all 50 ml for 1 gel (gel will be too thick – make 2 instead)
- Loading Dye: add glycerol solution to 25 mg bromophenol blue then add dH20 to 10 ml (to make 6X)
- Running gel: match wells to black side, run at 120 mA
- Visualize Gel in UV
- Turn power on
- Gel in machine face up
- Close door securely
- Turn white light on
- Adjust zoom, contrast, focus from black dial on top of machine
- Turn white light off (turns on UV)
- Press ‘live’ toggle – acq. Should be 0.4 sec.
- Print if desired or save on floppy disk
- Turn power off
- Dispose of gel in proper container
- Close door
May 25, 2009
- Made overnight bacteria culture (200 ml LB: 20 microliters DB3.1)
