Team:TorontoMaRSDiscovery/Notebook/May



=May 15, 2009=
 * 1) pH buffers received from VWR Mississauga
 * 2) *pH 4 buffer (red) 500 ml
 * 3) *pH 7 buffer (yellow) 500 ml
 * 4) *pH 10 buffer (blue) 500 mlAbove are used for pH/mV Meter calibration
 * 5) pH/mV meter calibrated according to manual – recorded in index
 * 6) Ethanol solution (70%) made from 85% ethanol

=May 19, 2009=
 * 1) 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
 * 1) 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
 * 1) 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=
 * 1) Retrieved autoclaved ddH20, glycerol solution
 * 2) Gel Electrophoresis (test run)
 * 3) *1% gel: 0.5 g agar mixed with 50 ml of 1X TBE
 * 4) *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)
 * 5) *Loading Dye: add glycerol solution to 25 mg bromophenol blue then add dH20 to 10 ml (to make 6X)
 * 6) *Running gel: match wells to black side, run at 120 mA
 * 7) Visualize Gel in UV
 * 8) Turn power on
 * 9) Gel in machine face up
 * 10) Close door securely
 * 11) Turn white light on
 * 12) Adjust zoom, contrast, focus from black dial on top of machine
 * 13) Turn white light off (turns on UV)
 * 14) Press ‘live’ toggle – acq. Should be 0.4 sec.
 * 15) Print if desired or save on floppy disk
 * 16) Turn power off
 * 17) Dispose of gel in proper container
 * 18) Close door

=May 25, 2009=
 * 1) Made overnight bacteria culture (200 ml LB: 20 microliters DB3.1)

=May 26, 2009=
 * 1) Took overnight cultures from incubator
 * 2) Inoculated 2 500 ml flasks with 25 ml of overnight culture (1 flask/culture)
 * 3) Placed 500 ml flasks into incubator at 37 degrees Celcius
 * 4) Grew overnights of DB3.1 from Waterloo (thanks :))