Descriptions

GSure Plasmid DNA Isolation Kit is an alkaline lysis based plasmid DNA isolation kit which employs silica membrane mini spin column for rapid and fast purification of the isolated DNA molecule. This kit can isolate covalently closed circular (ccc) plasmid DNA molecule from fresh overnight grown E.coli cells. Isolated plasmid is free from genomic DNA/Protein or RNA contamination. Isolation of plasmid DNA completes within 30 min. The kit can isolate at least 3μg plasmid DNA from 1ml of overnight grown culture of high copy number plasmid. For low copy number plasmid, yield generally varies from 500ng-1.5μg from 1ml culture. Yield could also be enhanced by chloramphenicol. GSure Plasmid DNA Isolation Kit can isolate plasmid DNA from 1ml to 12 ml of overnight grown cells. More than 75% of the isolated plasmid DNA is in ccc form which is the best suitable form of plasmid DNA molecule for any sort of downstream application. Most ccc form of plasmid DNA found upto 3ml of starting culture volume. More than 3ml culture does not inhibit plasmid DNA isolation, but the amount of ccc form of DNA decreases.

Gsure Plasmid DNA Isolation kit can isolate different sized plasmid DNA with equal efficiency. 2.6 kb plasmid to 18kb plasmid DNA molecule has been isolated by the kit efficiently. Isolated plasmid DNA is free of genomic DNA. A PCR reaction using 16S rDNA specific primer pair was set up on isolated plasmid DNA molecule, generated no amplification establishing that the plasmid DNA is free of trace amount of contaminating gDNA. Spectrophotometric absorbance data reveals that the isolated plasmid DNA is free of protein and RNA contamination. A260:A280 ratio varies between 1.79-1.83. Plasmid DNA isolated by GSure Plasmid DNA isolation kit is compatible for all sorts of downstream application like restriction digestion, PCR amplification, Sanger sequencing or in vitro transcription. GSure Plasmid DNA Isolation kit comes with a unique chromogenic indicator, sheerPink. SheerPink helps to ensure completion of lysis reaction properly. A complete lysis and proper neutralization assures ccc form of isolated plasmid and no genomic DNA contamination. GSure Plasmid DNA Isolation Kit contains 3 buffers which assure complete lysis of cells and digestion of contaminating proteins. Wash buffer supplied with the kit confirms complete removal of proteins and contaminants from the silica membrane. Optimized protocol guaranties extremely purified plasmid DNA isolation reproducibly. All the buffers should be stored at room temperature. Prescribed volume of absolute ethanol is to be added with the wash buffer before starting the work.

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Specification

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