Introduction

The MagPure Plasmid purification system uses the paramagnetic bead technology for high-throughput preparation of high-copy or low-copy plasmid DNA from E. coli cells. This kit also can be used with fosmid and BAC vector-based constructs. The system uses alkaline lysis followed by a MagPure purification to differentially bind plasmid DNA to paramagnetic beads. While the DNA is bound to the beads, contaminants can be rinsed away using a simple washing procedure. Because MagPure uses magnetic separation technology, the protocol does not require vacuum filtration. This makes kit extremely amenable to automation. Plasmid DNA purified with this system is most commonly used in Sanger Sequencing and PCR amplification.

Details

Specifications

Features	Specifications
Main Functions	Isolation up to 15μg endotoxin free plasmid DNA from 1-5ml bacterial culture
Applications	Enzyme digestion, sequencing, PCR and labeling,  etc.
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Sample type	Conventional plasmid, plasmid≤30KB
Sample amount	1-5ml
Elution volume	≥50μl
Time per run	≤80 minutes

Principle

This product is based on the purification method of high binding magnetic particles. The sample is lysed and digested under the action of lysate and Lysozyme. DNA is released into the lysate. After adding magnetic particles and binding solution, DNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption.The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally DNA was eluted by Elution Buffer.

Advantages

• High purity – purified plasmid can be directly used in sequencing, enzyme digestion and PCR, etc.
• Fast – it takes only 80 minutes to complete the isolation
• High yield – up to 15μg plasmid can be binded in one column
• Economic – excellent cost effectiveness performance

Kit Contents

Contents	P181402	P181403	P181404
Purification Times	100 Preps	500 Preps	5000 Preps
RNase A	10 mg	30 mg	2 x 160 mg
Buffer P1	30 ml	150 ml	2 x 800 ml
Buffer P2	30 ml	150 ml	2 x 800 ml
Buffer LEN3	20 ml	80 ml	800 ml
Buffer LN4	90 ml	400 ml	4 x 980 ml
MagPure Particles	3.5 ml	17 ml	3 x 60 ml

Storage and Stability

RNase A and MagPure Particles should be stored at 2–8°C upon arrival. However, short-termstorage (up to 12 weeks) at room temperature (15–25°C) does not affect its performance. Theremaining kit components can be stored dry at room temperature (15–25°C) and are stable for atleast 18 months under these conditions.The entire kit can be stored at 2–8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature whenused. If any precipitates form in the buffers,warm at 37℃ to dissolve. After addition of RNase A, Buffer P1 is stable for 6 months when stored at

The MagPure Plasmid purification system uses the paramagnetic bead technology for high-throughput preparation of high-copy or low-copy plasmid DNA from E. coli cells. This kit also can be used with fosmid and BAC vector-based constructs. The system uses alkaline lysis followed by a MagPure purification to differentially bind plasmid DNA to paramagnetic beads. While the DNA is bound to the beads, contaminants can be rinsed away using a simple washing procedure. Because MagPure uses magnetic separation technology, the protocol does not require vacuum filtration. This makes kit extremely amenable to automation. Plasmid DNA purified with this system is most commonly used in Sanger Sequencing and PCR amplification.

Overview

• Ensure optimal results for sensitive applications like NGS
• Up to a tenfold increase in microRNA mapping during sequencing runs to reduce costs
• Isolates all sizes of exosomal and extracellular vesicle RNA, including microRNA.
• Bind and elute all RNA irrespective of size or GC content, without bias.
• No phenol extractions.
• No Proteinase K treatment.
• No carrier RNA.
• Concentrate isolated RNA into a flexible elution volume ranging from 50 μL to 100 μL.
• Purify high-quality RNA in 15-20 minutes.
• Purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing.
• Purification is based on EXTRAClean spin column chromatography that uses Norgen’s proprietary separation matrix.

The kit is designed to isolate all sizes of RNA, including microRNA. The kit provides a clear advantage over other available kits in that it does not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or protease treatments. Moreover, the kit allows the user to elute into a flexible elution volume ranging from 50 μL to 100 μL. The purified RNA is of the highest integrity, and can be used in a number of downstream applications including real time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.

Details

Supporting Data

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Sample Type	Exosomes purified using Norgen’s Purification Kits
Size of RNA Purified	All sizes, including miRNA and small RNA (< 200 nt)
Elution Volume	50-100 μL
Time to Complete 10 Purifications	35-40 minutes
Average Yields	Variable depending on specimen

Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.