Our PCR and DNA Fragment Purification Kit is designed for rapid and efficient purification of DNA from PCR and other enzymatic reaction mixtures. The kit utilizes a proprietary silica-based membrane technology in the form of a convenient spin column, eliminating the need for tedious resin manipulations or toxic phenol-chloroform extractions. Our PCR and DNA Fragment Purification Kit effectively removes primers, dNTPs, unincorporated labeled nucleotides, enzymes and salts from PCR and other reaction mixtures. The kit can be used for purification of DNA fragments from 50 bp to 40 kb with recovery rates up to 100%. Each purification column has a total binding capacity of up to 30 μg of DNA and the entire procedure takes just 15 minuted. The purified DNA can be used in common downstream applications such as sequencing, restriction digestion, labeling, ligation, cloning, in vitro transcription, blotting or in situ hybridization.
Principle A reaction mixture containing DNA is combined with the binding buffer and added to a purification column. A chaotropic agent in the binding buffer denatures proteins and promotes DNA binding to the silica membrane in the column. As an added convenience, the binding buffer contains a color indicator that allows for easy monitoring of the solution pH for optimal DNA binding. Impurities are removed with a simple wash step. Purified DNA is then eluted from the column with the Elution Buffer. The recovered DNA is ready for use in downstream applications.
Features Highly efficient: 85-100% recoveries in the range of 50 bp-40 kb. Convenient: Spin columns are capped and assembled with collection tubes. Fast: Procedure takes 15 minutes. Pure DNA: A260/A280= 1.7-1.9.
Applications Fast and efficient purification of DNA fragments ideal for use in all conventional molecular biology procedures including:
conventional restriction digestion
automated fluorescent or radioactive sequencing
PCR
in vitro transcription
Quality Control The kit is tested in the purification of 50 bp and 1 kb PCR products according to the protocol. The quality of the purified DNA is evaluated spectrophotometrically by agarose gel electrophoresis, digestion with restriction enzymes and automated fluorescent sequencing.
Important Notes • Prior to the initial use of the kit, dilute the Wash Buffer(PE) with ethanol (100%):
Storage And Stability
PCR Purification Kit can be stored for up to 12 months at room temperature (15-25°C) or at 4°C for storage periods longer than 12 months. Any precipitate in the buffers can be re-dissolved by incubating at 37°C before use.
