Introduction

This product supplies a simple and rapid extraction of total RNA including microRNA from Blood, buffy coat, bone marrow, Cell suspension and other body fluids. The kit is based on superparamagnetic particles purification technology, no phenol-chloroform extraction or alcohol precipitation. Purified RNA is ready for downstream applications such as RT-PCR, virus RNA testing and so on.

Details

Principle

The Kit can be used for both manual extraction process and automatic nucleic acid extraction machines. This Kits is suitable for extracting RNA from ≤ 1 x10⁶ cells suspension, 50 μl Whole Blood, 50μl buffy coat, 20μl bone marrow. 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. RNA/DNA is released into the lysate. After adding magnetic particles and binding solution, DNA/RNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption. The adsorbed particles were digested using DNase and washed with washing buffer to remove proteins and impurities, washed with ethanol to remove salts, and

Kit Contents

Contents	R661101C	R661102C	R661103C
Purification times	48 Preps	96 Preps	480 preps
MagPure RNA Particles	1.0 ml	1.5 ml	6 ml
Proteinase K	24 mg	48 mg	220 mg
Protease Dissolve Buffer	1.8 ml	1.8 ml	15 ml
DNase I	600 μl	2 x 600 μl	10 x 600 µl
DNase Buffer	15 ml	15 ml	60 ml
Buffer AL	10 ml	10 ml	60 ml
Buffer MCB*	9 ml	9 ml	30 ml
Buffer MW1*	13 ml	22 ml	110 ml
Buffer MW2*	6 ml	20 ml	100 ml
RNase Free Water	10 ml	15 ml	120 ml

Storage and Stability

MagPure RNA Particles and Proteinase K should be stored at 2–8°C upon arrival. DNase I should be stored at -20°C. However, short-term storage (DNase I up to 1 weeks, MagPure RNA Particles and Proteinase K up to 8 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 stable for atleast 18 months under these conditions.

This product supplies a simple and rapid extraction of total RNA including microRNA from Blood, buffy coat, bone marrow, Cell suspension and other body fluids. The kit is based on superparamagnetic particles purification technology, no phenol-chloroform extraction or alcohol precipitation. Purified RNA is ready for downstream applications such as RT-PCR, virus RNA testing and so on.

The kit is developed for RNA quantification. The RNA Quantification HS kit includes HS Dye, HS Dilution Buffer, and two RNA Standards. Simply dilute the HS Dye with the HS Dilution Buffer, add RNA sample, then read the concentration using the Qubit Fluorometer. The assay is accurate for RNA concentrations from 250 pg/µL to 100 ng/µL (Figure 1). The RNA Quantification High Sensitivity Kit has several advantages over traditional RNA quantitation such as stability, sensitivity, and contaminant tolerance.

Features

• • • Optimized condition for using with the Qubit Fluorometer
    • Uses the Qubit RNA High Sensitivity assay setting
    • Cuts costs by 60%

The performance of the BioDynami RNA Quantification HS kit is nearly identical to that of Thermo Fisher’s Qubit RNA HS kit (figure below).

Common contaminants such as detergents, solvents, salts, free nucleotides, or protein are well tolerated in the assay (Table 1).

Qubit is a registered trademark of Thermo Fisher Scientific.

The kit is developed for RNA quantification. The RNA Quantification HS kit includes HS Dye, HS Dilution Buffer, and two RNA Standards. Simply dilute the HS Dye with the HS Dilution Buffer, add RNA sample, then read the concentration using the Qubit Fluorometer. The assay is accurate for RNA concentrations from 250 pg/µL to 100 ng/µL (Figure 1). The RNA Quantification High Sensitivity Kit has several advantages over traditional RNA quantitation such as stability, sensitivity, and contaminant tolerance.

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

Figure 1 / 6

Previous

Next

Click for expanded view

Kit Specifications
Minimum Urine Input	2 mL
Maximum Urine Input	10 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 the 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 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