Alberta NINT/24 July 2008


< Back to notebook

< Previous entry | Next entry >

lab work (SD):

            Measured A600 of cell cultures from 23/07/08 96-well plate.  Once samples were >0.3, 50 ul of cell
            culture from each well was transfered into 150 ul of MQ H20 in a clear 96-well plate.  The A600 was
            read.  It was very weak, and so the A600 for 150 ul of undiluted cell cultures was read.  To
            determine the "actual" A600 values read from the 96-well plate, 800 ul of 3 samples were measured
            for A600 in individual cuevettes.  Subsequent readings will have to be adjusted.
            1 ml Z buffer + 0.2 ul 10% SDS + 40 ul chloroform was aliquoted into a 96-well plate.  200 ul of
            cell culture was added to each well, pipetting up and down 15 times to lyse.  5 min was required to
            allow the chloroform to settle.
            150 ul of lysed cells was transfered into a clear 96-well plate and 150 ul of Z buffer was used as
            a blank control in the last column.
            20 ul ONPG was added to each well and the time of addition was recorded.
            No yellow color developed within one hour so the plate was left at 30 C overnight.