Revision as of 14:34, 28 June 2008

Friday 27 June

Chin Chong, Zhen Fu: Proceed with the characterization experiment for Quantification of RFU

Cells that were left to grow in the morning reached OD 1.2
Centrifuge the cells at 4400 rpm and at 25 deg C for 5 minutes
Remove the LB broth and Resuspend the cells with 5ml of M9 medium with glycerol
Incubate at 37 deg C and grow the cells to OD 1.2
Introduce the cells into the 96-well
Induce the cells with varying concentrations of lactose (1-10mM) in 1mM increments
Did Control 1: Cells with water, Control 2: M9 medium, Control 3: Water
Place the 96 well into FLx800™ Fluorescence Microplate Reader and followed the protocol as before and measure RFU for 12 hrs
Range of RFU interested was 10-70 RFU
Data points of 5 RFU increments ranging from 10 to 70 RFU were noted when the cella are allowed to grow for 12 hrs.
Whenever the desired RFU readings were measured, the Microplate Reader was paused and the desired well contents were extracted out into eppendrof tubes. For each desired RFU readings, two samples were extracted.
The tubes were centrifuged at 4400 rpm, 25 deg C for 5 minutes
The supernantant was removed carefully with a pippete so as to prevent removing any cell contents
The tubes containing only the cell contents were then stored in -80 deg C fridge for further experiment (SDS PAGE) in future

@@ Line 18: / Line 18: @@
 #The tubes were centrifuged at 4400 rpm, 25 deg C for 5 minutes
 #The supernantant was removed carefully with a pippete so as to prevent removing any cell contents
-#The tubes containing only the cell contents were then stored in -80 deg C fridge for further experiment in future
+#The tubes containing only the cell contents were then stored in -80 deg C fridge for further experiment (SDS PAGE) in future