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

GeneJET Plasmid Miniprep KitFast isolation ofhigh quality plasmid DNAP19

High Quality of Plasmid DNAThe GeneJET Plasmid Miniprep Kit is designed for fast and cost-effective isolation of high quality plasmid DNA fromrecombinant E.coli cultures. Up to 20µg of high copy plasmid DNA can be recovered from 1 to 5 ml of culture. Neitherphenol/chloroform extraction nor alcohol precipitation is required. The kit utilizes a silica-based membrane in the form ofa convenient spin column. The purifi ed plasmid DNA is ready for immediate use in all molecular biology procedures.Features• Efficient – yields up to 20µg of high quality plasmid DNA• Fast – procedure takes only 14 minutes• Easy-to-use – simple procedure, color-coded bottles• Convenient – GeneJET Spin Columns are capped and assembled with collection tubesGeneJET Spin Columns are cappedand assembled with collection tubesApplicationsFast and easy isolation of high quality plasmid DNA suitablefor all conventional molecular biology procedures, including:• FastDigest DNA cleavage• Conventional restriction digestion• PCR, fast PCR• Automated fl uorescent sequencing• Conventional radioactive sequencing• Transformation• In vitro transcription (see www.fermentas.com for recommendations)Kit Components• GeneJET Spin Columns assembled withCollection Tubes• Resuspension Solution• Lysis Solution• Neutralization Solution• Wash Solution (concentrated)• RNase A Solution• Elution Buffer

High Efficiency of ProcessEfficient & Reproducible Isolation of High Quality DNAGet a high yield of high quality plasmid DNA with GeneJET Plasmid Miniprep Kit – up to 20µg of high copy plasmidsand up to 10µg of low copy plasmids. The A 260/280 ratio of purifi ed DNA is in the range of 1.8-1.9. The actual plasmid yielddepends on the bacterial strain used for propagation, plasmid copy number and media used for cultivation.Higher DNA YieldConsistency of DNA Quality100RelaxedSupercoiledYield, %50M 1 2 3 4 5 6A pBluescript-based high copy number plasmid was purifi ed from E.coliculture with 6 different GeneJET Spin Columns.M – FastRuler DNA Ladder, High Range, ready-to-use (#SM1123)1-6 – plasmid DNA isolated with 6 different GeneJET Spin Columns0Fermentas Vendor 1 Vendor 2A pBluescript-based high copy number plasmid was purifi ed from 1.5mlof recombinant E.coli culture using the GeneJET Plasmid Miniprep Kitand miniprep purifi cation kits from other vendors. The total recoveredplasmid DNA yields were compared.GeneJET Plasmid Miniprep Kit guarantees reproducible and high quality results!Speed-Up DNA Cleavage with FastDigest Restriction EnzymesFermentas FastDigest restriction enzymes are specifi cally formulated for digestion of plasmid DNA in 5 minutes. Highquality plasmid DNA is required to achieve effi cient digestion. DNA purifi ed with the GeneJET Plasmid Miniprep Kit isideal for fast cleavage with FastDigest enzymes.Efficient DNA Cleavage in 5 MinutesA recombinant pJET1 plasmid containing 953bp insert was purifi ed from recombinant E.coliculture using the GeneJET Plasmid Miniprep Kit and digested in 5 minutes with FastDigest restriction enzymes.M 1 2 3 4 5 MM – GeneRuler Express DNA Ladder (#SM1551)1 – plasmid containing insert2 – plasmid containing insert, digested with FastDigest NotI (#ER0594)3 – plasmid containing insert, digested with FastDigest BamHI (#ER0054)4 – plasmid containing insert, double digested with FastDigest NotI and FastDigest BamHI5 – plasmid containing insert in the opposite orientation, double digested with FastDigest NotI and FastDigest BamHIVisit www.fermentas.com for detailed information about FastDigest restriction enzymesSpeed-up clone analysis with GeneJET Plasmid Miniprep Kit in combination withFastDigest restriction enzymes!www.fermentas.com

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