The EXTRACTME PLASMID MINI KIT is designed for a rapid and efficient purification of high quality plasmid DNA from recombinant Escherichia coli strains. The isolation protocol and buffer formulations were optimized for high isolation efficiency and DNA purity. The product is intended for research use only.
Bacterial broth culture, frozen cell pellet – however the efficiency will be decreased
Approx. 60 μg DNA
Approx. 25 minutes
A260/A280 ratio = 1.7 – 1.9
DNA purification procedure utilizes spin minicolumns with membranes which efficiently and selectively bind nucleic acids. During the first step plasmid DNA is released from bacterial cells by alkaline lysis. Then lysate is neutralized and all cell residues along with proteins and genomic DNA are separated in centrifugation step. Lysate is applied to purification minicolumn membrane and DNA is bound. A two-step washing stage effectively removes impurities and enzyme inhibitors. Purified plasmid DNA is eluted with then use of a low ionic strength buffer (Elution Buffer) or water (pH 7.0–9.0) and may be used directly in all downstream applications such as PCR, qPCR, Southern blotting, DNA sequencing, enzymatic restriction, ligation and so forth, or stored until ready to use.
BalticBlue is an indicatior that provides visual identification of optimum buffer mixing. This prevents common handling errors that lead to inefficient cell lysis and incomplete precipitation of SDS, genomic DNA and cell debris. BalticBlue should be added to Resuspension Buffer at a ratio of 1:1000 (e.g. 12.5 μl BalticBlue to 12.5 ml Resuspension Buffer). Mix thoroughly. BalticBlue precipitates.
After addition of Lysis Buffer, precipitate dissolves and color changes to blue. During neutralization invert mixture gently until white precipitate is formed and blue solution will be colorless. When isolating in parts, transfer enough of Resuspension Buffer for your isolations to a separate bottle/tube and add BalticBlue.
The quality of each production batch (LOT) of the EXTRACTME PLASMID MINI KIT is tested using standard QC procedures. Purified DNA concentration and quality are evaluated by gel electrophoresis and spectrophotometer.
Manufacturer: BLIRT S.A.