Clontech
login | register | order now | view cart | feedback
Products
Nucleic Acid Purification


Products >  Nucleic_Acid_Purification >  Plasmid >  Miniprep-Transfection-grade >  Spin

A fast way to purify plasmids for transfection—NucleoSpin Plasmid Transfection-grade

NucleoSpin Plasmid Transfection-grade is a fast miniprep kit that uses a new technology developed by Macherey-Nagel to reduce the level of endotoxins co-purified during plasmid preparations from bacterial lysates. Since endotoxins interfere with eukaryotic cell survival, endotoxin reduction is essential prior to cell transfection.

Principle/procedure

The NucleoSpin Plasmid Transfection-grade procedure is a modified version of the Birnboim and Doly alkaline lysis plasmid miniprep protocol. Pelleted bacteria are resuspended in Buffer A1 and plasmid DNA is liberated from the cells by SDS/alkaline lysis with Buffer A2. Buffer A3 neutralizes the lysate; precipitates genomic DNA, proteins, and cell debris; and creates appropriate conditions for binding of plasmid DNA to the silica membrane.

The crude lysate is cleared by centrifugation and brought into contact with a silica membrane which binds plasmid DNA on its surface. Endotoxins and proteins are removed by the innovative Detoxification Buffer ERB. Further contaminants such as salts are removed with ethanolic Buffer AQ, while traces of ethanol are removed by centrifugation.

Pure plasmid DNA is eluted under low ionic strength conditions with slightly alkaline Buffer AE (5 mM Tris/HCl, pH 8.5) and is ready for any common downstream application, including transfection (research use only).

  At-A-Glance   Documents   Images & Data

Features

  • Novel technology to diminish endotoxin content (<50 EU/µg DNA) for successful transfection
  • A miniprep purification format simplifies your workflow
  • Ultra-fast procedure

Applications

  • Typical downstream applications: transfection of common cell lines (lipofection, electroporation)
  • Also suitable for standard molecular biology applications such as cloning and sequencing

Product Overview

Technology Silica membrane and
endotoxin-reduction technologies
Format Miniprep spin columns
Endotoxin Content <50 EU/µg DNA
(EU=Endotoxin Units)
Starting Material Up to 5 ml bacterial culture
(E. coli, high-copy plasmids)
Vector Size <15 kb
Typical Yield 5–20 µg
A260/280 1.80–1.85
Elution Volume 30–50 µl
Preparation Time 14 min/6 preps
Binding Capacity 25 µg

 
 
Products
New Offers Cat. # Product Contents Size Price Units Select
740490.50 NucleoSpin® Plasmid Transfection-grade 50 Preps $83.00
740490.250 NucleoSpin® Plasmid Transfection-grade 250 Preps $348.00
 

 

 
 

Takara Bio USA is a Takara Bio Company © 2017 Privacy Policy | Terms & Conditions | Terms of Use