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Life Science > Molecular Biology > DNA & RNA Purification > Five-Minute Miniprep Kit
DNA & RNA Purification

Five-Minute Miniprep Kit


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.


5 Minute Prep Kit Testimonials  
5_minute_Overnight_Protocol

Beneficial Features
  • Use only 400 µl overnight bacterial culture

  • Purify up to 5 µg from high-copy plasmid

  • Only 5 minutes from start to finish

  • Choose vacuum or spin protocols

Order Information
Product Product Description Technical
Bulletin		 Preps
PFM10 GenElute Five-Minute Plasmid Miniprep Kit Technical_Bulletin_icon 10
PFM50 GenElute Five-Minute Plasmid Miniprep Kit Technical_Bulletin_icon 50
PFM250 GenElute Five-Minute Plasmid Miniprep Kit Technical_Bulletin_icon 250

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Sample Data

Purified Plasmid DNA is Especially Suited for Automated Sequencing

Figure 1. Capillary DNA sequencing of plasmid DNA purified with the GenElute Five-Minute Plasmid Miniprep Kit. pCMV-SPORT-βgal (7.9 kb) was prepared from 400 µl overnight culture of E. coli 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.

Click Image to View Sequence in its Entirety

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Achieve Consistent Yields and Quality for Common Applications

Figure 2. Restriction digestion analysis of pCMV-SPORT-βgal DNA purified with GenElute Five-Minute Plasmid Miniprep Kit. Lanes 1 & 19: 1 kb DNA ladder (Sigma D 0428); lanes 2 to 17: 16 independent plasmid mini preparations; lane 18: uncut pCMV-SPORT-βgal DNA. Each digestion was carried out in 15 µl at 37 °C for 1 hour, with 3 µl of eluate and 3 units each of Eco RI and Xho I. A quarter of each digestion was fractionated in 1% agarose gel.

agarose gel image for 5 minute miniprep

*Sigma Advanced Technology™ distinguishes the scientific team and the product it developed for the achievement of a new technology, a novel invention or measurable improvements resulting in absolute performance advantages over current methods.

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