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Reagents / Solutions Lysis Buffer: 10ml 10% Sodium dodecyl sulphate (SDS) 10ml Glycerol 10ml b-mercaptoethanol 8ml 0.5M Tris pH6.8 1ml 0.1% bromophenol blue 51ml H2O Protocol Spin down 106 cells and wash, if required, in phosphate buffered saline. Resuspend cells in 100µl warmed lysis buffer, vortex then boil for 5 - 10 minutes to break up DNA. Cool (not on ice - the SDS will precipitate) and centrifuge to collect droplets. Vortex briefly to mix. The samples may now be analysed by , or Store samples at -20℃ until required. |
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