Overview

The E.Z.N.A.® HP Plasmid Mini Kit I is designed to isolate up to 30 µg of high-quality plasmid DNA from 1-5 mL bacterial cultures from strains that are endonuclease positive. The HP Plasmid DNA Kit includes a protease digestion step to remove endonucleases and proteins from the sample. Plasmid DNA purification is simplified with HiBind® Mini Column technology into three quick steps: bind, wash and elute. Purified plasmid DNA is immediately ready for a wide variety of downstream applications such as routine screening, restriction enzyme digestion, and manual/automated fluorescent DNA sequencing.

• Rapid – Purification of plasmid DNA in 30 minutes or less
• Safe – No phenol/chloroform extractions
• Versatile – Spin and vacuum formats available
• High-quality – DNA is suitable for downstream applications

Specifications

Product Data

DNA Concentration Comparison

Figure 1.   pGEM plasmid was purified from 4 mL DH5α cultures harboring the plasmid and eluted in 50 µL volume using kits from Omega Bio-tek and Company Q according to manufacturer’s recommended protocols. Plasmid DNA concentration was determined by optical density measurements using Thermo Scientific’s NanoDrop™ 2000c system. 

Super-Coiled DNA

Figure 2.   5 µL of purified pGEM plasmid was analyzed on a 1% Agarose gel. Plasmid was isolated from 1 mL DH5α cultures harboring the plasmid using Omega Bio-tek’s E.Z.N.A.® Plasmid Mini Kit I. 

DNA yield and quality using E.Z.N.A.® Plasmid Mini Kit I

Table 1.  pGEM plasmid was purified from 1 mL DH5α cultures harboring the plasmid and eluted in 50 µL volume. Plasmid DNA concentration was determined by optical density measurements using Thermo Scientific’s NanoDrop™ 2000c system.
 

Endotoxin Level of Purified Plasmid DNA using E.Z.N.A.® Plasmid Mini Kit I

Table 2.  Plasmid DNA purified with E.Z.N.A. Plasmid Mini Kit I was used in a 5 µL Sanger sequencing reaction.  DNA was analyzed on an Applied Biosystems 3730XL. 

Endotoxin Level of Purified Plasmid DNA using E.Z.N.A.® Plasmid Mini Kit I

Table 3.  Endotoxins in plasmid DNA preps.  Plasmid DNA was isolated from 0.8 mL LB cultures following each manufacturer’s recommended protocols. Endotoxin levels were determined with Thermo Scientific’s Pierce LAL Chromogenic Endotoxin Quantitation Kit. 

자료보기