Manual

Product Number: PLK1401

Shipping and Storage

1. When using for the first time, add all RNase A carried by the reagent kit to Buffer P1 (final concentration 100μg/ml) and store at 4℃. If RNase A is inactivated in Buffer P1, the extracted plasmid may contain trace amounts of RNA residue. Adding RNase A to Buffer P1 is sufficient.
2. Buffer ER can be transported at room temperature and stored at 4℃ for one month. It can be stored for a long time at -20℃.
3. When the ambient temperature is low, SDS in Buffer P2 may precipitate and become turbid or precipitate. It can be heated in a 37℃ water bath for a few minutes to restore clarity. Do not shake violently to avoid excessive foam formation.
4. To avoid volatilization, oxidation, and pH changes caused by prolonged exposure of reagents to the air, each solution should be covered tightly in a timely manner after use.

Components

Description

This reagent kit extracts plasmid DNA from cultured bacteria using alkaline lysis method. It uses a unique solution formula and endotoxin clearance reagent, and only requires a few simple centrifugations to remove impurities such as proteins, polysaccharides, endotoxins, RNA, etc., to obtain high-quality plasmid DNA. The OD260/280 of purified DNA is usually around 1.8, and the resulting plasmid can be directly applied in tasks that require high DNA purity, such as cell transfection and even animal in vivo experiments. The purification process in the later stage is operated in a 1.5ml small centrifuge tube, which is simple, does not require special equipment, does not require column passing, and does not require phenol chloroform extraction; Plasmids released by bacterial lysis can be completely recovered without worrying about the loss of plasmid DNA. This method extracts and purifies plasmid DNA with minimal damage to plasmids. Even large plasmids or ultra large BAC/PAC plasmids with a size of 10kb or even 100kb can be effectively purified as long as they can be extracted by alkaline lysis. You can choose to dissolve plasmids in any small volume, with a concentration of up to 5μg/μl. The super helix ratio can reach up to 95%, with no endotoxins and good transfection effect.

Features

1. No need to use toxic reagents such as phenol and chloroform, and no need for ethanol precipitation. Fast and convenient, 0.5-2mg of pure high copy plasmid DNA can be quickly extracted from 150-200ml of Escherichia coli LB (Luria Bertani) culture medium, with an extraction rate of 80-90%.
2. The obtained plasmid yield is high, with a super helix ratio of up to 95% and a concentration of up to 5μg/μl. Good purity, can be directly used for various molecular biology experiments such as enzyme digestion, transformation, PCR, in vitro transcription, sequencing, etc.
3. The endotoxin content is extremely low (<0.1EU/gDNA) and can be directly applied to cell transfection.

Application

Suitable for the preparation of large amounts of high-purity or transfection grade plasmids and the preparation of large BAC/PAC plasmids

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