DescriptionHighly efficient purification of plasmid DNA by combining fast SDS/alkaline lysis of the starting material with the speed and purity of silica-membrane column purification.Product HighlightsFast - streamlined protocol for extraction of eighteen plasmid samples in as little as 15 minutesEfficient – rapid SDS/alkaline lysis and efficient column-binding for high-yield recovery of all plasmid moleculesHigh-performance – efficient extraction of high-purity plasmid DNA, ideal for use in all downstream applicationsFlexible – reliable recovery of high- and low-copy plasmids from all gram-positive bacteriaProduct DescriptionThe ISOLATE II Plasmid Mini Kit provides a simple, efficient column-based method for the isolation of plasmid DNA from bacterial cultures, without the need for hazardous reagents such as phenol.By combining SDS/alkaline lysis with the speed and ease-of-use of silica-membrane purification, the ISOLATE II Plasmid Mini Kit provides a fast method for the purification of high-quality plasmid DNA. Separate protocols are provided for the isolation of high-copy plasmids, the isolation of low-copy plasmid, P1 constructs and cosmid DNA from E. coli and the isolation of plasmids from gram-positive bacteria such as Bacillus, Staphylococcus, Bifidobacteria or Corynebacteria.The ISOLATE II Plasmid Mini Kit has been designed to deliver optimal performance in cloning, sequencing as well as end-point PCR alongside any enzyme from the Meridian PCR portfolio, including MyTaq DNA Polymerase. Additionally, the ISOLATE II Plasmid Mini Kit can be used in tandem with the SensiFAST Real-Time PCR Kits for high-performance qPCR.Request InfoApplicationsCloningSequencingRestriction digestionLabelingEnd-point PCRqPCRTransfectionFluorescent sequencingIn vitro transcription Reproducible resultspUC19 plasmid was isolated from 5 mL E. coli overnight LB cultures using the ISOLATE II Plasmid Mini Kit and cut with various restriction enzymes, before analysis on 1% TAE agarose gel. Each restriction digestion (lanes 1-6) was performed in triplicate on plasmids from different overnight cultures. The results illustrate consistency in plasmid recovery. High plasmid recoverypUC19 plasmid was isolated from increasing volumes of E. coli from an overnight LB culture (0.05 ml, 0.1 mL, 1 mL, 3 mL and 5 mL culture, lanes 1- 5 respectively), using ISOLATE II Plasmid Mini Kit and run on 1.5% TAE agarose gel. The results illustrate the very high binding capacity of the columns in the ISOLATE II Plasmid Mini Kit. Excellent performance of isolated plasmid DNA in downstream applicationspET19 plasmid was isolated from 3 mL E. coli overnight LB culture using ISOLATE II Plasmid Mini Kit. Sequencing was carried out based on standard T7 promoter and terminator primers, allowing more than 750 nucleotides to be read.Nucleic Acid Isolation Guide Download the ISOLATE II Guide with detailed product descriptions and performance data to help you choose the best product for your research ISOLATE II Selection Chart