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Nucleic Acid Purification


Products >  Nucleic_Acid_Purification >  Plasmid >  Miniprep >  Spin

Miniprep—NucleoSpin Plasmid

The NucleoSpin Plasmid Miniprep Kit is designed for the rapid (25 min), high-yield (up to 40 µg), small-scale preparation of high-purity plasmid DNA. These miniprep columns contain a unique silica membrane that binds up to 40 µg of plasmid DNA. The purified DNA is suitable for transfection, cloning, sequencing, PCR, transformation, and restriction analysis.

The miniprep kit allows purification of up to 25 μg plasmid DNA per preparation, from 1–5 ml of saturated E. coli culture. You can also use this kit to purify up to 40 μg of plasmid DNA from 5–10 ml E. coli culture. In order to obtain optimal results from 5–10 ml E. coli culture, increase your buffer volumes using the NucleoSpin Buffer Set.

Using the NucleoSpin Plasmid Miniprep Kit

Harvested bacteria are resuspended and processed by SDS/alkaline lysis (see the User Manual, located in the Documents tab below). High-salt buffer is added to neutralize the lysate and to create appropriate conditions for DNA binding to the silica membrane. After centrifugation, the clear supernatant is loaded onto a NucleoSpin Plasmid miniprep spin column.

Contaminants like salts and macromolecular cellular components are removed by simple washing with ethanolic Buffer A4. Additional washing with Buffer AW is recommended for the following demands:

  • Complete removal of high levels of endonucleases, with buffer AW prewarmed to 50°C (e.g. from wild-type E. coli strains)
  • Processing 5–10 ml of bacterial culture
  • Long read length in DNA sequencing
  • Better performance of critical enzymatic reactions

Highly pure plasmid miniprep DNA is finally eluted in 50 μl slightly alkaline Buffer AE (5 mM Tris/HCl, pH 8.5). Alternatively, water (pH 8.0–8.5) can be used.

At-A-Glance Documents Images & Data Resources

Features

  • Ready-to-use plasmid DNA—18 minipreps in 25 min
  • Obtain up to 40 µg of high-purity miniprep plasmid DNA
  • Silica membrane technology—no resin or phenol extraction
  • Reproducible yields

Applications

  • Transfection
  • Cloning
  • Sequncing
  • PCR
  • Transformation
  • Ligation
  • Restriction analysis

Product Overview

Technology Silica membrane
Format Miniprep spin columns
Starting Material 1–5 ml (standard protocol)
5–10 ml (Requires increased buffer volumes;
purchase the NucleoSpin Buffer Set)
Vector Size <15 kb
Typical Yield <25 µg (from 1–5 ml starting material)
<40 µg (from 5–10 ml starting material)
A260/280 1.70–1.85
Elution Volume 50 µg
Preparation Time 25 min/18 preps

 
 
Products
New Cat. # Product Package Size Price # of Units Select
  740588.250 NucleoSpin® Plasmid 250 Preps $279.00
  740588.50 NucleoSpin® Plasmid 50 Preps $67.00
740499.240C NucleoSpin® Plasmid 240 Preps $279.00
  740499.50 NucleoSpin® Plasmid (No Lid) 50 Preps $67.00
  740499.250 NucleoSpin® Plasmid (No Lid) 250 Preps $279.00
 

 

Recommended Products
Cat. # Product Package Size Price # of Units Select
740953 NucleoSpin® Plasmid Buffer Set Each $39.00
740588.50S NucleoSpin® Plasmid Binding Columns 50 Columns $34.00
740588.250S NucleoSpin® Plasmid Binding Columns 250 Columns $140.00
740499.50S NucleoSpin® Plasmid Binding Columns (No Lid) 50 Columns $34.00
740499.250S NucleoSpin® Plasmid Binding Columns (No Lid) 250 Columns $140.00
 
 


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