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Description: The GenElute™ Five-Minute Miniprep Kit features an ultra streamlined protocol yielding up to 5 µg high-quality plasmid DNA in about five minutes.
- Sophisticated – no pelleting cells or clearing lysates
- Flexible – binding column works with any standard laboratory vacuum manifold
- Plentiful – up to 5 µg of plasmid DNA or enough for 15 sequencing reactions
Scientists at Sigma developed a novel lysis procedure that eliminates pelleting cells, removing media and clearing neutralized lysates. This discovery enables super-rapid purification of plasmid DNA, and qualifies the product for the Sigma Advanced Technology* designation.
Procedure: 400 µl of culture is added to a 2 ml collection tube. Cells are lysed for 1-2 minutes. A Binding Solution is added and the mixture is passed through a binding column. After washing the column to remove trace contaminants, the plasmid DNA is eluted in 10mM Tris HCl, pH 8.5. Molecular biology water may be substituted as an elution reagent.
Application: The DNA is suitable for immediate use in capillary DNA sequencing, clone screening, restriction digestion and PCR. |
 was prepared from 400 µl overnight culture of <i>E. coli</i> host strain DH5α with the GenElute Five-Minute Plasmid Miniprep Kit. A 5 µl sample of the preparation was submitted, with adjustment in DNA concentration, to SeqWright DNA Technology Service (Houston, TX) for sequencing with a T7 sequencing primer and ABI BigDye™ terminator v3.1 on an ABI Prism® 3730 DNA Analyzer. The Phred > 20 score was 905 bases.</p>');)
