Overview

• Concentrate circulating DNA into a flexible elution volume ranging from (25 µL – 50 µL)
• Isolate inhibitor-free cell-free circulating DNA
• Compatible with Streck Cell-Free DNA BCT® Tubes
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix
• Research use format is also available

These kits provide a fast, reliable and convenient spin column method for the isolation of high quality, high purity and inhibitor-free cell-free circulating DNA (cfc-DNA) from plasma/serum sample. These kits are designed to isolate all sizes of cfc-DNA from either fresh or frozen plasma/serum samples and the purified DNA is eluted into a flexible elution volume ranging from 25 µL to 50 µL. The purified plasma/serum cfc-DNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive PCR and Southern Blot analysis, microarrays and NGS.

Background

Plasma/Serum cell-free circulating DNA (cfc-DNA) has the potential to provide biomarkers for certain cancers and disease states as well as fetal DNA in maternal blood. Currently, significant advancements are being made in utilizing cfc-DNA as biomarkers for the early diagnosis, prognosis and monitoring of therapy for several cancer types and autoimmune diseases. Cell-free mitocondrial DNA (cfmtDNA) is also under investigation for its clinical significance. This cfc-DNA is usually present as short fragments of less than 1000 bp. In addition, cell-free fetal DNA has been widely used as a non-invasive method for prenatal diagnosis including early identification of fetal sex, genetic studies for families at high risk for inherited genetic disorders, screening for Rhesus factor, screening for aneuploidy and identification of preeclampsia.

Plasma/Serum Cell-Free Circulating DNA Purification Micro Kit Dx

For serum input volumes 10 µL – 200 µL. Purify high-quality DNA in 15-20 minutes

Plasma/Serum Cell-Free Circulating DNA Purification Mini Kit Dx

For serum input volumes 200 µL – 500 µL. Purify high-quality DNA in 15-20 minutes.

Plasma/Serum Cell-Free Circulating DNA Purification Midi Kit Dx

The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 25 µL to 50 µL. All components for the purification & concentration are provided in one convenient & fast kit for the easy processing of large input volumes of bodily fluids. For a schematic workflow of the protocol click here to view. For serum input volumes 1 mL – 4 mL. Purify high-quality DNA in 90 minutes.

Plasma/Serum Cell-Free Circulating DNA Purification Maxi Kit Dx

The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 25 µL to 50 µL. All components for the purification & concentration are provided in one convenient & fast kit for the easy processing of large input volumes of bodily fluids. For a schematic workflow of the protocol click here to view. For serum input volumes 5 mL – 10 mL. Two kits in one – purify and concentrate DNA from bodily fluids using a convenient two column system.

Details

Supporting Data

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Kit Specifications
Minimum Plasma/Serum Input	10 μL
Maximum Plasma/Serum Input	200 μL
Size of DNA Purified	≥ 50 bp
Elution Volume	25-50 μL
Time to Complete 10 Purifications	15-20 minutes
Average Yields*	Variable depending on specimen

*Please check page 7 in the product manual for the Plasma/Serum Average Yields and the Common DNA Quantification Methods.

Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature (15-25°C) for up to 2 years without showing any reduction in performance. Norgen’s Plasma/Serum Cell-Free Circulating DNA Purification Kits contain ready-to-use Proteinase K solution, which is dissolved in a specially prepared storage buffer. The Proteinase K is stable for up to 2.5 years after delivery when stored at room temperature. To prolong the lifetime of Proteinase K, storage at 2–8°C is recommended.

Component	Cat. Dx55100 (50 preps)	Cat. Dx55500 (50 preps)	Cat. Dx55600 (20 preps)	Cat. Dx55800 (10 preps)
Binding Buffer B	2 x 40 mL	40 mL	2 x 85 mL 1 x 12 mL	2 x 85 mL 1 x 40 mL
Proteinase K	2 x 1 mL	0.6 mL	6.5 mL	8 mL
Solution WN	18 mL	18 mL	18 mL	18 mL
Wash Solution A	38 mL	18 mL	18 mL	18 mL
Elution Buffer B	8 mL	8 mL	30 mL	30 mL
Micro Spin Columns	50	50	20	10
Mini Spin Columns	50	–	–	–
Midi Spin Columns	–	–	20	–
Maxi Spin Columns	–	–	–	10
Collection Tubes	100	50	20	10
Elution Tubes (1.7 mL)	100	50	20	10
Product Insert	1	1	1	1

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 20µg plasmid DNA from 1-3ml 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	1-3ml
Elution volume	≥50μl

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

1. Suitable for extracting plasmids from 1-5ml or <3ml YT medium.
2. The same amount of buffer 1, 2, and 3 avoids errors caused by adjusting the pipette, making it convenient to use in conjunction with automated workstations.
3. Containing buffer 1 for washing, reducing the problem of false high production.
4. The purified plasmid can be directly used for sequencing, enzyme digestion, PCR, and other applications.

Kit Contents

Contents	P181102	P181103	P181104
Purification Times	100 Preps	500 Preps	5000 Preps
RNase A	10 mg	50 mg	2 x 250 mg
Buffer P1	30 ml	150 ml	2 x 750 ml
Buffer P2	30 ml	150 ml	2 x 750 ml
Buffer N3	30 ml	150 ml	2 x 750 ml
Buffer PW1	35 ml	180 ml	2 x 900 ml
MagPure Particle NB*	2.2 ml	11 ml	2 x 60 ml

Storage and Stability

RNase A and MagPure Particle NB should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect its performance. The remaining kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. After addition of RNase A, Buffer P1 is stable for 6 months when stored at 2-8°C.

Experiment Data

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.