Fast Midiprep Plasmid DNA Purification Kit

Introduction

Plasmids are circular, extrachromosomal DNA molecules that act as carriers, or vectors, for a specific DNA segment in molecular biology. Bacteria are employed to reproduce plasmids, allowing your desired DNA to be mass-produced. The method by which researchers acquire plasmids from bacteria is known as plasmid purification.

Purifying plasmid DNA from bacterial cells is a critical step in cloning. Bacterial cells are lysed during plasmid purification, which liberates DNA and other biological components from the cell wall. After removing cellular components, the DNA-containing lysate is treated to eliminate impurities and separate the plasmid DNA from the genomic DNA.

The plasmid DNA extracted is the same quality as that produced by purification with two passes through cesium chloride gradients—the most stringent approach for DNA purification. High-quality DNA can be purified in less than twenty minutes for transfections and other applications. There is no need for extra processes to remove impurities such as RNA, proteins, or endotoxins. Furthermore, the protocol’s exclusion of phenol, chloroform, ethidium bromide, and cesium chloride reduces exposure to and disposal of hazardous compounds.

Principle & Workflow

Hygia Pure Midiprep Plasmid DNA Purification uses weak anion exchange beads to quickly purify plasmid DNA. The beads allow for plasmid DNA binding directly. This approach uses typical cell resuspension, alkaline lysis, and neutralization processes and allows plasmid DNA to bind directly to the beads. A single bead wash removes impurities such as RNA, proteins, metabolites, and other low molecular weight molecules. Unique wash buffers ensure that salts, proteins, RNA, and other biological components are eliminated, allowing concentrated, highly pure DNA to be eluted. The ultrapure plasmid DNA is subsequently eluted with a high-salt buffer. The DNA is desalted, concentrated with an isopropanol precipitation process, and centrifuged. The entire treatment takes roughly twenty minutes to complete.

Feature and Benefits

● Simple protocols- no columns or filters required.

● Quick results- purify plasmid DNA in less than twenty minutes.

● No Phenol/chloroform extractions

● Low endotoxin

● Quality – DNA is suitable for most downstream applications

● Scalable – Easily adjusts for sample size and automation

● Reproducible results