Introduction

The Kit is specially designed for simultaneous purification of genomic DNA and total RNA from formalin-fixed, paraffin-embedded (FFPE) tissue sections. Purified DNA/RNA is suitable for use in applications such as real-time PCR and pyrosequencing.

Details

Specifications

Features	Specifications
Main Functions	Co-isolation DNA and RNA from FFPE tissue
Applications	RT-PCR, cDNA synthesis, PCR and second-generation sequencing, etc.
Purification method	Polydisperse silicon based magnetic beads (DNA)Monodisperse carbonyl magnetic beads (RNA)
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Adaptive instrument	Nucleic acid extractor and pipetting workstation
Sample type	FFPE slice, FFPE puncture sample, embedded tissue
Sample amount	No more than six 10 µm sections of 150 mm2 surface area or three 20µm sections of 150 mm2 surface area
Yield	DNA: 1 – 10 μg,  RNA: 1 – 25 μg

Principle

The sample is lysed and digested under the action of lysate and protease. DNA/RNA is released into the lysate. After adding magnetic particles and binding solution, DNA will be adsorbed on the surface of magnetic particles, the supernatant contain RNA was collected and bind to MagBind Particles. The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally DNA/RNA was eluted by Elution Buffer.

Advantages

• High quality – high purity total RNA / DNA can be directly used in various downstream applications
• Safe – no phenol chloroform extraction required
• Simultaneous extraction – simultaneously isolate DNA and RNA from one sample
• Post digestion sorting – higher DNA and RNA yields
• High RNA yield – monodisperse carbonyl adsorption

Kit Contents

Contents	R632701	R632702	R632703
Purification Times	48 Preps	96 Preps	5 x 96 Preps
MagBind Particles	1.1 ml	2.5 ml	11 ml
MagPure Particles N	1.1 ml	2.5 ml	11 ml
Proteinase K	24 mg	48 mg	220 mg
Protease Dissolve Buffer	3 ml	10 ml	15 ml
Buffer DPS	50 ml	100 ml	2 x 250 ml
Buffer ATL	20 ml	30 ml	120 ml
Buffer BST1	20 ml	40 ml	200 ml
Buffer BXW1*	44 ml	110 ml	3 x 110 ml
RNase Free Water	15 ml	30 ml	120 ml

Storage and Stability

Proteinase K, MagPure Particles N and MagBind Particles should be stored at 2–8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15–25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15–25°C) and are stablefor at least 18 months under these conditions.

Experiment Data

The Kit is specially designed for simultaneous purification of genomic DNA and total RNA from formalin-fixed, paraffin-embedded (FFPE) tissue sections. Purified DNA/RNA is suitable for use in applications such as real-time PCR and pyrosequencing.

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
• The purified exosomal RNA is free from any circulating RNA-binding proteins
• No phenol extractions, Proteinase K treatment, nor carrier RNA
• No time-consuming ultracentrifugation, filtration nor special syringes are required
• No precipitation reagents, nor overnight incubation required
• Concentrate isolated exosomal RNA and are 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 plasma/serum 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 as well as all sizes of the free-circulating protein-bound RNA, including microRNA. These kits provide a clear advantage over other available kits in that they do not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or protease treatments. Moreover, these 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.

Plasma/Serum Exosome and Free-Circulating RNA Isolation Mini Kit

For sample volumes ranging from 50 µL to 1 mL.

Plasma/Serum Exosome and Free-Circulating RNA Isolation Midi Kit

For sample volumes ranging from 1 mL to 4 mL.

Plasma/Serum Exosome and Free-Circulating RNA Isolation Maxi Kit

For sample volumes ranging from 4 mL to 10 mL.

Details

Supporting Data

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Kit Specifications
Minimum Plasma Input	1 mL
Maximum Plasma Input	4 mL
Size of RNA Purified	All sizes, including miRNA and small RNA (< 200 nt)
Elution Volume	50 – 100 µL
Time to Complete 10 Purifications	40 – 45 minutes
Average Yields*	Variable depending on specimen

*Please check page 5 of the product insert for the average yields and the common RNA quantification methods

Storage Conditions and Product Stability
All solutions 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
This kit is suitable for the purification of exosomes from fresh or frozen serum or plasma prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used as heparin can significantly interfere with many downstream applications such as RT-PCR.

Component	Cat. 59500 (50 preps)	Cat. 59600 (25 preps)	Cat. 59700 (15 preps)
Slurry E	12.5 mL	12.5 mL	12.5 mL
ExoC Buffer	8 mL	8 mL	2 x 8 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

DBCO-PEG12-Maleimide is a PEG linker containing a DBCO moiety and a terminal primary maleimide group. The maleimide group will react with a thiol group to form a covalent bond, enabling the connection of biomolecule with a thiol. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

DBCO-PEG12-Maleimide is a PEG linker containing a DBCO moiety and a terminal primary maleimide group. The maleimide group will react with a thiol group to form a covalent bond, enabling the connection of biomolecule with a thiol. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.