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


Products >  Nucleic_Acid_Purification >  Plasmid >  Miniprep-Transfection-grade >  96-Well

A fast way to purify plasmids for transfection in 96-well plates—NucleoSpin 96 Plasmid Transfection-grade

The NucleoSpin 96 Plasmid Transfection-grade kit uses a new technology developed by Macherey-Nagel to reduce the level of endotoxins co-purified during plasmid preparations from bacterial lysates, in a 96-well-plate format. Since endotoxins interfere with eukaryotic cell survival, endotoxin reduction is essential prior to cell transfection.

Principle/procedure

The NucleoSpin 96 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).

Required hardware for vacuum processing and centrifugation

The NucleoSpin 96 Plasmid Transfection-grade kits can be used manually with the NucleoVac 96 Vacuum Manifold. Additionally, a suitable centrifuge for harvesting the bacteria (plate or tube centrifuge) is required.

  At-A-Glance   Documents   Images & Data

Features

  • Novel technology to diminish endotoxin content (<50 EU/µg DNA) for successful transfection
  • Innovative MN Wash Plate minimizes risk of cross-contamination
  • Ultra-fast procedure
  • NucleoSpin 96 Plasmid Transfection-grade Core Kits provide basic content focusing on automation platforms. (Additional accessories can be combined as needed.)

Applications

  • Manual and automated isolation of high-copy plasmids
  • Typical downstream applications: transfection of common cell lines, cell-based assays, screenings

Product Overview

Technology Silica membrane and
endotoxin-reduction technologies
Format 96-well plate
Processing Manual or automated, vacuum
or centrifugation
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 100–200 µl
Preparation Time 45 min/plate
Binding Capacity 35 µg

 
 
Products
New Cat. # Product Contents Size Price Units Select
740491.1 NucleoSpin® 96 Plasmid Transfection-grade 96 Preps $289.00
740491.4 NucleoSpin® 96 Plasmid Transfection-grade 4 x 96 Preps $1,029.00
740491.24 NucleoSpin® 96 Plasmid Transfection-grade 24 x 96 Preps $5,119.00
740492.4 NucleoSpin® 96 Plasmid Transfection-grade Core Kit 4 x 96 Preps $899.00
740492.24 NucleoSpin® 96 Plasmid Transfection-grade Core Kit 24 x 96 Preps $4,490.00
 

 

 
 

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