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Cat. No.: PMPMB-200 (for 200T)

Cat. No.: PMPMB-1k (for 1000T)

Description

Plasmid Mini Preparation Kit with Magnetic Beads is a reagent kit that utilizes a novel nucleic acid purification medium coated with magnetic beads, designed for the stable, efficient, and convenient extraction of small quantities of plasmids from Escherichia coli.

The plasmids extracted using this kit can be directly used for experiments such as cell transfection, DNA sequencing, PCR, PCR-based mutagenesis, in vitro transcription, bacterial transformation, and restriction enzyme digestion.

Plasmids are fully released using an alkaline lysis method, and then they specifically bind to the magnetic beads. Under the influence of an external magnetic field (such as a magnetic separation rack), the magnetic beads can be rapidly and efficiently separated from the corresponding solution. After thorough washing to remove impurities, the plasmids are finally eluted from the magnetic beads using elution buffer, resulting in high-quality plasmid samples.

Components

• Magnetic Beads
• Solution I (Suspension Buffer)
• Solution II (Lysis Buffer)
• Solution III (Binding Buffer)
• Solution IV (Wash Buffer)
• Solution V (Elution Buffer)
• RNase A (100mg/ml)

Features

• This kit offers several advantages, including stable extraction results, high purity, and flexible and convenient operation. The plasmid extraction system of this kit has been repeatedly tested and optimized, yielding approximately 10-15µg of high-copy plasmids from 2-3ml overnight cultures of Escherichia coli. Plasmid extraction can be completed in as fast as approximately 20 minutes. Following standard operating procedures, this kit is suitable for plasmid extraction from 1-5ml overnight bacterial cultures. Due to the flexible adjustment of the amount of magnetic beads used, if extracting medium-copy plasmids, you can simply scale up the extraction system proportionally, for example, by doubling it.
• Currently, the primary method for plasmid extraction is column purification, which typically involves repetitive centrifugation or the use of specialized filtration devices. Moreover, the fiber cutting that occurs during column purification can have some impact on the supercoiled state of plasmids. In contrast, the magnetic bead method operates under mild conditions, eliminating the need for cumbersome centrifugation or filtration steps and replacing them with a simple magnetic adsorption process, ensuring a quick and convenient operation.
• Compared to traditional extraction methods, this kit does not involve the use of toxic reagents like phenol/chloroform during the extraction process.

Storage

Store at room temperature, effective for one year. When Magnetic Beads are not in use for an extended period, they can be stored at 4°C, which allows for even longer storage.

Note

• You will need to provide anhydrous ethanol and a magnetic separation apparatus.
• Before the first use, add all the provided RNase A to Solution I (Suspension Buffer), mix well, and label the bottle. Store at 4°C after adding RNase A.
• Before the first use, add the specified amount of anhydrous ethanol to each bottle of Solution IV (Wash Buffer), mix well, and label the bottle.
• Magnetic beads may settle when left undisturbed. Before use, ensure adequate mixing by gently vortexing or inverting several times until well mixed.
• Before magnetic separation, gently shake the centrifuge tube to disperse the magnetic beads fully before bringing them close to the magnetic field. If bead aggregation occurs, gently agitate the liquid inside the tube to allow the adhered beads to flow down.
• Please use the recommended bacterial culture volume. If the culture volume is too large, it may lead to magnetic bead aggregation, which can affect washing and subsequently impact plasmid purity. In case of bead aggregation, disperse the beads as much as possible during washing to improve extraction efficiency. If bead aggregation occurs, it is advisable to reduce the bacterial culture volume in subsequent experiments.
• At lower temperatures, Solution II (Lysis Buffer) and Solution III (Binding Buffer) may produce precipitates. Check before use. If there is precipitation, heat at 37°C in a water bath, mix well, and then use.
• Do not mix Solution II (Lysis Buffer) too vigorously as it may produce a large number of bubbles. After use, ensure the bottle cap is tightly sealed to prevent carbonation from CO2 in the air.
• This product is suitable for manual extraction and can also be used with workstations or nucleic acid extraction instruments.
• The yield of extracted plasmid DNA may vary depending on factors such as plasmid copy number. The OD value of extracted plasmid DNA may also slightly fluctuate due to different bacterial strains.
• Solution II (Lysis Buffer), Solution III (Binding Buffer), and Solution IV may have irritant effects on the human body. Handle them with care and take appropriate precautions.
• This product is for research use by professionals only and should not be used for clinical diagnosis or treatment. It should not be used in food or drugs and should not be stored in residential homes.
• For your safety and health, wear lab attire and disposable gloves while handling the product.

Only for research and not intended for treatment of humans or animals

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