Introduction

Hipure viral RNA kit is suitable for purifying viral RNA from samples such as cell-free body fluid or culture medium. The kit is based on silica gel column purification technology. It requires no toxic phenol chloroform extraction and time-consuming alcohol precipitation in the extraction. The whole extraction process takes only 25 minutes. The kit is suitable for extracting viral RNA from 1-140µl cell-free fluid samples such as serum, plasma, cell-free body fluid or culture medium. The product has successfully extracted hepatitis B A/C, hepatitis C RNA, SARS and HIV. The obtained RNA can be directly used for RT-PCR, Northern hybridization and virus detection.

Details

Specifications

Features	Specifications
Main Functions	Extract viral RNA from 140μl cell-free samples
Applications	RT-PCR, Northern hybridization, and various virus detection
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Cell-free body fluid or culture medium
Sample amount	140μl
Elution volume	≥15μ
Time per run	≤25 minutes
Liquid carrying volume per column	800μl
Binding yield of column	100μg

Principle

Hipure silica gel column is based on glass fiber filter membrane with high binding force. Under the condition of high concentration of ionizing agent (such as guanidine hydrochloride or guanidine isothiocyanate), the filter membrane can adsorb nucleic acid through hydrogen bond and electrostatic, while protein and other impurities are not adsorbed and removed. The filter membrane adsorbed with nucleic acid is washed to remove the residual protein and salt. Finally, the nucleic acid adsorbed on the filter membrane can be washed out with low salt buffer (such as buffer TE) or water. The obtained nucleic acid has high purity and can be directly used in various downstream experiments.

The sample is homogenized and lysed in the lysate, and RNA is released into the lysate. The high concentration of guanidine isothiocyanate contained in the lysate denatured and inactivated endogenous or exogenous RNase, while RNA is protected from degradation. The lysate is centrifuged to remove insoluble impurities. After adding ethanol to adjust the binding conditions, it is transferred to the column for filtration. RNA is adsorbed on the membrane of the column, while protein is removed without adsorption. The column is washed with buffer VHB to remove protein and other impurities, washed with buffer RW2 to remove salt. Finally the RNA is eluted by RNase free water. The eluted RNA can be directly used in RT-PCR, Northern blot, poly-A purification, in vitro translation, etc.

Advantages

• Fast – several samples can be extracted in 20 minutes by column method
• High quality – high purity total RNA can be directly used in various sensitive downstream applications
• Safe – no phenol chloroform extraction required
• Sensitive – RNA can be recovered at the level of PG
• High yield – carrier RNA contained in the product maximize the recovery of trace nucleic acid

Kit Contents

Contents	R417102	R417103
Purification Times	50 Preps	250 Preps
HiPure Viral Micro Columns	50	250
2ml Collection Tubes	100	500
Buffer VRL	50 ml	200 ml
Carrier RNA	310 µg	3 x 310 µg
Buffer VHB*	13 ml	110 ml
Buffer RW2*	20 ml	2 x 50 ml
Nuclease Free Water	10 ml	30 ml

Storage and Stability

Carrier RNA 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 stable for at least 18 months under theseconditions

Hipure viral RNA kit is suitable for purifying viral RNA from samples such as cell-free body fluid or culture medium. The kit is based on silica gel column purification technology. It requires no toxic phenol chloroform extraction and time-consuming alcohol precipitation in the extraction. The whole extraction process takes only 25 minutes. The kit is suitable for extracting viral RNA from 1-140µl cell-free fluid samples such as serum, plasma, cell-free body fluid or culture medium. The product has successfully extracted hepatitis B A/C, hepatitis C RNA, SARS and HIV. The obtained RNA can be directly used for RT-PCR, Northern hybridization and virus detection.

Overview

• Fractionate leukocytes from whole blood in minutes with provided RBC Lysis Buffer
• Isolate total RNA, including microRNA, without phenol
• Rapid and convenient spin-column format and 96-well plate for high throughput applications
• Purified RNA is ready for any downstream application including RT-PCR, qRT-PCR, NGS, arrays and more. Find out more information on Norgen’s NGS services
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

These kits provide a rapid method for the isolation and purification of total leukocyte (white blood cell) RNA from mammalian blood samples. These kits are supplied with an RBC (red blood cell) Lysis Buffer for selective removal of red blood cells and fractionation of leukocytes by centrifugation. Isolation of leukocyte RNA results in improved expression profiling and other downstream applications by removing the masking effects of some RNAs which are very abundant in whole blood, such as globin mRNAs. These kits are able to isolate total leukocyte RNA, including both large mRNA and all small RNA species containing microRNA (miRNA) and small silencing RNA (siRNA). The purified RNA is of the highest quality and can be used in a number of downstream applications.

Leukocyte RNA Purification Kit (Spin Column)

This kit provides a rapid method for the isolation and purification of total leukocyte RNA from mammalian blood samples in 40 minutes. Allowable blood input ranges from 10 μL to 2 mL or 3 x 10⁶ Leukocytes

Leukocyte RNA Purification Plus Kit (Plus)

Norgen’s Leukocyte RNA Purification Plus Kit provides a rapid method for the isolation and purification of total leukocyte (white blood cell) RNA from up to 3 mL of mammalian blood samples. Complete 10 purifications in as little as 40 minutes.

Leukocyte RNA Purification 96-Well Kit (High Throughput)

Purification is based on 96-well column chromatography using Norgen’s proprietary resin as the separation matrix. Purification can be performed using either a vacuum manifold or centrifugation. Norgen’s kit allows for the isolation of total leukocyte RNA, including all small RNA species. The purified RNA is of the highest quality 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. Allowable blood input ranges from 10 μL to 1 mL or 1.5 x 10⁶ Leukocytes.

Details

Supporting Data

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Kit Specifications
Maximum Column Binding Capacity	Up to 50 μg RNA
Maximum Loading Volume	650 μL
Size of RNA Purified	All sizes, including small RNA <200 nt
Minimum Blood Input	10 μL
Maximum Blood Input	2 mL or 3 x 106 leukocytes
Time to Complete 10 Purifications	40 minutes
Input Type	Blood from Human, Hamster, Mouse, Rabbit, and Rat
Average Yield Human Blood (500 μL) Hamster Blood (100 μL)	1.5 μg 2.5 μg

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. The RBC Lysis Buffer should be stored at 4°C upon arrival.

Component	Cat. 21200 (50 preps)	Cat. 21250 (50 preps)	Cat. 37800 (2 x 96 preps)
RBC Lysis Buffer	2 x 100 mL	2 x 1 L	2 x 90 mL
Buffer RL	30 mL	30 mL	–
Binding Solution	–	–	2 x 40 mL
Wash Solution A	38 mL	38 mL	–
Wash Solution	–	–	2 x 30 mL
Elution Solution A	6 mL	6 mL	–
Elution Solution	–	–	2 x 20 mL
Enzyme Incubation Buffer	–	6 mL	1
DNase I	–	1 Vial	–
Mini Spin Columns	50	–	–
Lysate Homogenization Column	–	50	–
Single Cell RNA Column	–	50	–
RBC Lysis 96-Well Plate	–	–	2
96-Well Filter Plate	–	–	2
Adhesive Tape	–	–	4
Collection Tubes	50	100	–
96-Well Collection Plate	–	–	2
Elution Tubes (1.7 mL)	50	50	–
96-Well Elution Plate	–	–	2
Product Insert	1	1	1

Sulfo DBCO-PEG3-acid is an analog of DBCO-Acid with PEG linker and a DBCO group. The DBCO groups is commonly used for copper-free Click Chemistry reactions due to its strain promoted high energy. The hydrophilic PEG chain and sulfo group increase water solubility. The terminal carboxylic acid can react with primary amine groups in the presence of activators (e.g. EDC, or HATU) to form a stable amide bond. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Sulfo DBCO-PEG3-acid is an analog of DBCO-Acid with PEG linker and a DBCO group. The DBCO groups is commonly used for copper-free Click Chemistry reactions due to its strain promoted high energy. The hydrophilic PEG chain and sulfo group increase water solubility. The terminal carboxylic acid can react with primary amine groups in the presence of activators (e.g. EDC, or HATU) to form a stable amide bond. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.