Home  Contacts  Order  Catalog  Support
 Search  Alphabetical Index  Numerical Index
 Restriction Enzymes  Modifying Enzymes  PCR, qPCR, RT-PCR & dNTPs  Molecular Cloning  Nucleic Acid Purification
 Molecular Labeling & Detection  In vitro Transcription  Electrophoresis Products  Nucleotides  Transfection Reagents  Reagents
C A T A L O G
 

GeneJET™ Plasmid Miniprep Kit

#K0502 for 50 preps 
#K0503 for 250 preps

Related Documents (in pdf, ~50-400 KB):

Detailed Protocol: #K0502, #K0503
MSDS (English)
MSDS (English-USA)
MSDS (German)
Brochure
Flyer
QuickProtocol™

 

Features


 brochure in pdf, 426 KB


flyer in pdf, 220 KB

Description
 The GeneJET™ Plasmid Miniprep Kit provides a simple, rapid and cost-effective method for the isolation of high quality plasmid DNA from recombinant E.coli cultures. The 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.

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 conventional and fast digestion with restriction enzymes, PCR, transformation and automated sequencing.

Applications
 Fast and easy isolation of high quality plasmid DNA suitable for all conventional molecular biology procedures, including:
- FastDigest® restriction digestion;
- conventional restriction digestion;
- PCR, fast PCR;
- automated fluorescent sequencing;
- conventional radioactive sequencing;
- transformation;
- in vitro transcription (see Recommendations below).

Quality Control
 The kit is tested for the isolation of pUC19 DNA from E.coli according to the protocol. The quality of isolated DNA is evaluated by spectrophotometric assay, agarose gel electrophoresis, digestion with restriction enzymes and automated fluorescent sequencing.

 
Figure 1. Overview of purification procedure.

Components of the Kit

Recommendations for obtaining high quality RNA transcripts
  • Use DEPC-treated Water and DEPC-treated (RNase-free) tubes for plasmid DNA elution.
  • Do not add RNase A solution to Resuspension Solution, if the purified plasmids are intended for use in in vitro transcription.
  • If RNase A was used during purification, phenol/chloroform extraction of plasmid DNA before or after plasmid digestion is recommended.
    (Phenol extraction should be followed by chloroform extraction (3 times) and precipitation with NaCl in 2.5 volumes of ethanol).

Figure 2. 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 (see the protocol for fast digestion and analysis).
M1, M2 - ZipRuler™ Express DNA Ladder Set (#SM1373)
C - control pUC19 DNA digested with FastDigest® EcoRI and FastDigest® HindIII
1-6 - miniprep DNA from recombinant clones, double digested with FastDigest® EcoRI and FastDigest® HindIII


Figure 3. Electropherogram demonstrates the high quality sequencing data of plasmid DNA purified with the GeneJET™ Plasmid Miniprep Kit.
A pUC19-based plasmid was purified using the GeneJET™ Plasmid Miniprep Kit from E.coli DH10B. The sequencing reaction was run on an ALFexpress™ II sequencer.

 Home  Search  Contacts  Order  Catalog  Support

Updated kovo 04, 2008 11:19