GeneJET Plasmid Miniprep Kit

Available On-Site

Room Temperature

The GeneJET Plasmid Miniprep Kit is a simple, rapid, and cost-effective system for the isolation of high quality plasmid DNA from recombinant E. coli cultures.
  
Loading...

The Thermo Scientific GeneJET Plasmid Miniprep Kit utilizes an exclusive silica-based membrane technology in the form of a convenient spin column. The kit recovers up to 20 µg of high copy plasmid DNA per isolation procedure.

Highlights

  • Efficient – high yields of up to 20 µg of high quality plasmid DNA
  • Fast – procedure takes less than 14 minutes
  • Convenient – no phenol/chloroform extraction or alcohol precipitation required
  • Pure – purified DNA is immediately ready to use

Applications

  • Fast isolation of high purity plasmid DNA suitable for all conventional molecular biology procedures, including:
    • FastDigest or conventional restriction digestion
    • Automated fluorescent and radioactive sequencing
    • PCR
    • In vitro transcription
    • Transformation

Principle

A bacterial culture is harvested and lysed. The lysate is then cleared by centrifugation and applied on the silica column to selectively bind DNA molecules at a high salt concentration. The adsorbed DNA is washed to remove contaminants, and the pure plasmid DNA is eluted in a small volume of elution buffer or water. The purified DNA is ready for immediate use in all molecular biology procedures, such as fast and conventional digestion with restriction enzymes, PCR, in vitro transcription, transformation, and automated sequencing.

Components

  • Resuspension Solution
  • Lysis Solution
  • Neutralization Solution
  • Wash Solution (concentrated)
  • RNase A Solution
  • Elution Buffer
  • Spin Columns
  • Collection Tubes
  • Detailed Protocol
  
HazardousNo
Quality Control
  • The kit is tested for the isolation of pUC19 DNA from E. coli according to the protocol.
  • The quality of the isolated DNA is evaluated by spectrophotometric assay, agarose gel electrophoresis, digestion with FastDigest restriction enzymes, and automated sequencing.
Storage ConditionAmbient,-20 C
Overview of purification procedure

Overview of purification procedure

Overview of purification procedure


 Fast clone analysis

Fast clone analysis

 Fast clone analysis

  • A 2.3 kb PCR fragment was cloned into pUC19 vector. Plasmid DNA from overnight bacterial cultures of recombinant clones was purified using the GeneJET Plasmid Miniprep Kit and analyzed by double digestion with FastDigest EcoRI and FastDigest HindIII in 5 minutes at 37°C.
  • M1, M2 – ZipRuler Express DNA Ladder Set
  • C – control pUC19 DNA digested with FastDigest EcoRI and HindIII
  • 1-6 – miniprep DNA from recombinant clones, double digested with FastDigest EcoRI and HindIII


Electropherogram demonstrates the high quality sequencing data of plasmid DNA purified with GeneJET Plasmid Miniprep Kit

Electropherogram demonstrates the high quality sequencing data of plasmid DNA purified with GeneJET Plasmid Miniprep Kit

Electropherogram demonstrates the high quality sequencing data of plasmid DNA purified with GeneJET Plasmid Miniprep Kit

Plasmid DNA was purified using the GeneJET Plasmid Miniprep Kit from E. coli and sequenced with a T7 sequencing primer. The sequencing reaction was analyzed on an ABI® 3730 DNA Analyzer. More than 750 nucleotides can be read.