Introduction

This product provides fast and easy methods for purification of total DNA for reliable PCR and southern blotting. Total DNA(e.g., genomic, viral, mitochondrial) can be purified from whole blood, plasma, serum, buffy coat, bone marrow, other body fluids, lymphocytes, cultured cells.

Relevant reagents were pre-packed in 96-well plates in accordance with the optimal protocol.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from 200μl whole blood
Applications	PCR, southern bolt and virus detection, etc
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Sample type	Anticoagulant blood, concentrated blood, buffy coat, lymphocytes and cultured cells
Sample amount	200μl
Elution volume	≥50μl
Time per run	≤60 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 Protease. 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 binding force – suitable for handling DNA rich samples, such as whole blood, buffy coat, concentrated blood, etc.
• Fast – polydisperse magnetic beads, fast magnetic response and short extraction time
• High purity – the obtained DNA can be directly used for second-generation sequencing, PCR based detection, gene bank, etc.
• Automatic – saving time andlabor and safer

Kit Contents

Contents	D631101	D631102	D631103
Purification Times	48	96	480
MagPure Particles	1.2 ml	2.5 ml	11 ml
Proteinase K	24 mg	50 mg	220 mg
Protease Dissolve Buffer	1.8 ml	5 ml	15 ml
Buffer AL	15 ml	30 ml	120 ml
Buffer BD	5 ml	10 ml	50 ml
Buffer GW1	22 ml	53 ml	220 ml
Elution Buffer	15 ml	30 ml	100 ml

Storage and Stability

Proteinase K, MagPure Particles 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 their 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.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 when used.

This product provides fast and easy methods for purification of total DNA for reliable PCR and southern blotting. Total DNA(e.g., genomic, viral, mitochondrial) can be purified from whole blood, plasma, serum, buffy coat, bone marrow, other body fluids, lymphocytes, cultured cells.

Relevant reagents were pre-packed in 96-well plates in accordance with the optimal protocol.

DBCO-C2-alcohol is a linker containing a DBCO moiety and a terminal primary hydroxyl group. DBCO group can react with azides in copper-free Click Chemistry reactions. The hydroxyl can react with a variety of functional groups. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

DBCO-C2-alcohol is a linker containing a DBCO moiety and a terminal primary hydroxyl group. DBCO group can react with azides in copper-free Click Chemistry reactions. The hydroxyl can react with a variety of functional groups. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

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Overview

• Isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias
• Versatile sample input ranges
• Isolate all sizes of free-circulating RNA, including microRNA​
• Bind and elute all RNA irrespective of size or GC content, without bias
• The purified exosomal RNA is free from any circulating RNA-binding proteins
• No phenol extractions, Proteinase K treatment nor carrier RNA required
• No time-consuming ultracentrifugation, filtration nor special syringes are required
• No precipitation reagents nor overnight incubation required
• Concentrate isolated exosomal RNA and free-circulating RNA into a flexible elution volume ranging from 50 µL to 100 µL
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

These kits provide a fast, reliable and convenient method to sequentially isolate and concentrate exosomal RNA as well as Free-Circulating RNA from different urine sample volumes. The purification is based on spin column chromatography that employs Norgen’s proprietary resin. These kits are designed to isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias. These kits provide a clear advantage over other available kits since they do not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or protease treatments. Moreover, the kits allow the user to elute into a flexible elution volume ranging from 50 µL to 100 µL. The RNA isolated from the purified exosomes is free from any protein-bound circulating RNA and is of the highest integrity. Moreover, the free-circulating, protein-bound, RNA is free from any exosomal RNA. The purified RNA 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.

Urine Exosome and Free-Circulating RNA Isolation Mini Kit

For sample volumes ranging from 250 µL to 1 mL.

Urine Exosome and Free-Circulating RNA Isolation Midi Kit

For sample volumes ranging from 2 mL to 10 mL.

Urine Exosome and Free-Circulating RNA Isolation Maxi Kit

For sample volumes ranging from 11 mL to 30 mL.

Details

Supporting Data

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Kit Specifications
Minimum Urine Input	250 µL
Maximum Urine Input	1 mL
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

*Please check page 4 of the product insert for the average yields and the common RNA quantification methods.

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.

Important Note
Urine samples stored at -70°C, -20°C or at 4°C will develop some precipitation due to the aggregation of some of the highly abundant proteins in urine. Eliminating these precipitates using centrifugation or filtration may cause the loss of exosomes. Furthermore, these precipitates may affect the quality of the purified nucleic acid. We recommend the use of Norgen’s Urine Preservative when collecting urine samples, which is designed for the preservation of nucleic acids and proteins in fresh urine samples at ambient temperatures. The components of the Urine Preservative allow samples to be stored for over 2 years at room temperature with no detected degradation of urine DNA, RNA or proteins. Norgen’s Urine Preservative is available as a liquid format in Norgen’s Urine Preservative Single Dose Ampules, as well as in a dried format in Norgen’s Urine Collection and Preservation Tubes.

Component	Cat. 59200 (50 preps)	Cat. 59300 (25 preps)	Cat. 59400 (15 preps)
Slurry E	12.5 mL	12.5 mL	12.5 mL
ExoC Buffer	8 mL	30 mL	50 mL
ExoR Buffer	12 mL	12 mL	12 mL
Lysis Buffer A	2 x 20 mL	2 x 20 mL	2 x 20 mL
Lysis Additive B	2 mL	2 mL	2 mL
Wash Solution A	2 x 18 mL	18 mL	18 mL
Elution Solution A	2 x 6 mL	6 mL	6 mL
Mini Filter Spin Columns	50	25	15
Mini Spin Columns	100	50	30
Collection Tubes	100	50	30
Elution Tubes (1.7 mL)	100	50	30
Product Insert	1	1	1