Team:Paris/3 August 2009

NoteBook
August 3rd

 

Brain work
Meeting this morning for the lab work week. Discuss about the schedule algorithm for the iPhone application.

 

Lab work
DID dying for membrane fluorescence microscopy ->try another method : FM 4-64 Miniprep digestion by XbaI and PstI Gel Migration Culture ON for Miniprep  
 * wash cell with MgSO4 then dying and incubation 30 min 25°C ->observation : 1/100 fluorescent cell and some vesicles ?!
 * dying in medium, incubation 30min 37°C ->observation : no significant (1/100)
 * dying in medium, incubation 30min 37°C, wash with MgSO4 ->observation : no significant (1/100)
 * For P1 pSB2K3, P2 BBa_J61002, P3 BBa_J32015, P4 BBa_B0014, P5 BBa_I14032, P7 BBa_R0040, P8 BBa_I0500.
 * Digestion Mix
 * 50µl Miniprep
 * 2µl XbaI
 * 2µl PstI
 * 0,6µl BSA
 * 6µl Buffer n°2
 * 2h at 37°C
 * Verification by migration on 1% Agarose gel with BET
 * 5µl of digestion solution
 * 1µl Loading (LD)
 * [P1|P2|P3|P4|P5|Ladder 100bp|P7|P8]
 * P1 = pSB2K3 = plasmid backbone = 4399bp (expected weight)
 * P2 = BBa_J61002 = plasmid backbone = (will not be digest by this enzyme)
 * P3 = BBa_J32015 = pelB = 66bp
 * P4 = BBa_B0014 = Double terminater = 95bp
 * P5 = BBa_I14032 = pLac = 37bp
 * P7 = BBa_R0040 = tetR repressible promoter = 54bp (weak but present, cf other photos)
 * P8 = BBa_I0500 = pBad/araC = 1210bp
 * S7 DH5&alpha;(g3p)