Brochure "GeneJET™ Plasmid Miniprep Kit" (© 2006 Fermentas)

Fast ProcedurePurified DNA is Ready To Use for Automated Fluorescent SequencingA pUC19-based plasmid was purifi ed from E.coli culture usingthe GeneJET Plasmid Miniprep Kit. An 400ng aliquot ofpurifi ed DNA was sequenced using the CycleReader AutoDNA Sequencing Kit (#K1721) with Cy5 labeled M13/pUCsequencing primer. The sequencing reaction was run on anALFexpress II sequencer. 780 bases have been read in thisrun using the plasmid purifi ed with the GeneJET PlasmidMiniprep Kit as a template.Overview of Purification ProcedureQuickProtocol GeneJET Plasmid Miniprep KitNote. All steps should be carried out at room temperature.All centrifugations should be carried out in a microcentrifuge at 12000 x g (~11000rpm).12Resuspend Cells, Lyse and NeutralizeAdd to the pelleted cells:250µl of Resuspension Solution (with RNase A) and vortex.250µl of Lysis Solution and invert the tube 4-6 times.350µl of Neutralization Solution and invert the tube 4-6 times.Centrifuge 5min.Bind DNALoad the supernatant to GeneJET spin column.Centrifuge 1min.3Wash the columnAdd 500µl of Wash Solution and centrifuge for 30-60s.Discard the flow-through. } Repeat twiceCentrifuge empty column for 1min.4Elute purified DNAAdd 50µl of Elution Buffer to the column and incubate 2min.Centrifuge 2min.Collect the flow-through.QuickProtocol and GeneJET are Fermentas trademarkswww.fermentas.com© 2006 Fermentas QP06Product Catalog # Size, µl#K0501 25 prepsGeneJET Plasmid Miniprep Kit#K0502 50 preps#K0503 250 prepsFastDigest restriction enzymes see www.fermentas.com for detailed informationCycleReader, FastDigest, FastRuler, GeneJET, GeneRuler and QuickProtocol are Fermentas trademarks.ALFexpress is Amersham Pharmacia Biotech trademark.Fermentas – a member of your research teamwww.fermentas.com© Fermentas 2006