Overview

• Extract total RNA (including microRNA) from FFPE samples
• No phenol extraction step
• Includes DNase for optional on-column DNA removal
• Isolated RNA is of the highest quality and integrity
• Isolate a diversity of RNA species
• Purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

Norgen’s FFPE RNA Purification Kits provide a rapid method for the isolation and purification of total RNA (including microRNA) from formalin-fixed paraffin-embedded (FFPE) tissue samples in as little as 1 hour. Using formalin to fix tissues leads to crosslinking of the RNA and proteins, and the process of embedding the tissue samples can also lead to fragmentation of the RNA over time. Norgen’s FFPE RNA Purification Kits provide conditions that allow for the partial reversing of the formalin modifications, resulting in a high quality and yield of RNA. These kits are able to purify all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA), depending on the age of the FFPE tissue as fragmentation of the RNA is known to occur over time. The RNA is preferentially purified from other cellular components without the use of phenol or chloroform.

FFPE RNA Purification Kit (Spin Column)

Maximum loading volume of 650 μL per column, and a maximum binding capacity of 50 μg per column.

FFPE 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. Maximum loading volume of 400 μL per well, and a maximum binding capacity of 50 μg per well.

Details

Supporting Data

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Kit Specifications
Maximum Binding Capacity Per Well	50 μg
Maximum Loading Volume Per Well	400 μL
Size of RNA Purified	All sizes, including small RNA (< 200 nt)
Maximum Amount of Starting Material:	5 slices of < 20 µm thick paraffin  25 mg of unsectioned block

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. The DNAse I should be stored at -20°C upon arrival. The Proteinase K should be stored at -20°C upon arrival and after reconstitution. This kit is stable for 1 year after the date of shipment.

Component	Cat. 25300 (50 preps)	Cat. 25400 (2 x 96 preps)
Digestion Buffer A	25 mL	2 x 25 mL
Buffer RL	30 mL	2 x 30 mL
Enzyme Incubation Buffer	6 mL	2 x 6 mL
Wash Solution A	38 mL	2 x 38 mL
Elution Solution A	6 mL	2 x 20 mL
Proteinase K	12 mg	2 x 20 mg
DNase I	1 vial	2 x 500μL
Micro Spin Columns	50	–
96-Well Incubation Plate	–	2
96-Well Plate	–	2
Adhesive Tape	–	8
Collection Tubes	50	–
96-Well Collection Plate	–	2
Elution Tubes (1.7 mL)	50	–
96-Well Elution Plate	–	2
Product Insert	1	1

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.

Description

Nucleic acid testing (NAT) is the method of choice for detection and quantification of a wide range of micro organisms. Primerdesign manufactures and supplies high quality quantitative real-time PCR kits for the detection and simultaneous quantification of numerous significant pathogens . A copy number standard curve is provided for quantification and an the internal extraction template (DNA or RNA), controls for the quality of the nucleic acid extraction and eliminates false negative results.

The kit is designed with the broadest possible detection profile to ensure that all clinically relevant strains and subtypes are detected. Target sequences are selected by working with data from key opinion leaders in the field. Multiple sequence alignments and unprecedented real-time PCR expertise in design and validation ensure the best possible kit. Details of the target and priming specificity are included in the individual handbooks above.

Packaged, optimised and ready to use. Expect Better Data.

Exceptional value for money
Rapid detection of all clinically relevant subtypes
Positive copy number standard curve for quantification
Highly specific detection profile
High priming efficiency
Broad dynamic detection range (>6 logs)
Sensitive to < 100 copies of target Accurate controls to confirm findings 150 reactions