This product is suitable for extracting total pathogen RNA from biological samples with no/low cell content such as body fluid, serum, plasma, soaking solution, tissue homogenate supernatant, culture medium supernatant, etc. The purified RNA can be used for clinical in vitro detection.

It is special designed for Pathogen RNA extraction from Sputum samples, can be used in tuberculosis (TB) detection.

Details

Specifications

Features	Specifications
Main Function	Extract total pathogen RNA from cell-free/low-content cell biological samples such as body fluids, serums, plasma, tissue homogenate supernatant, special design for sputum samples. Used in tuberculosis detection.
Applications	RT-PCR，PCR
Products	Pathogen RNA
Purification method	Polydisperse magnetic beads
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Sample type	cell-free/low-content cell biological samples such as body fluids, serums, plasma, tissue homogenate supernatant, sputum
Sample amount	200-300μl
Adaptive instrument	Nucleic acid extractor, pipetting workstation

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. 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 washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally DNA/RNA was eluted by Buffer NFW.

Advantages

• Fast – several samples can be extracted in 40 minutes by column method
• High quality – high purity total RNA / DNA can be directly used in various sensitive downstream applications
• Safe – no phenol chloroform extraction required
• Sensitive – DNA/RNA can be recovered at the level of PG

Kit Contents

Contents	IVD6672C
Purification Times	200 Preps
2ml Bead Tube	4 x 50
MagPure Particle	7.0 ml
Proteinase K	100 mg
Protease Dissolve Buffer	6 ml
DTT （Powder)	2g
Buffer SDS	15 ml
Buffer MLBN	220 ml
DNase I	4 x 0.6 ml
DNase Buffer	60 ml
Buffer MW1*	53 ml
Buffer MW2*	50 ml
Buffer NFW	30 ml

Storage and Stability

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

This product is suitable for extracting total pathogen RNA from biological samples with no/low cell content such as body fluid, serum, plasma, soaking solution, tissue homogenate supernatant, culture medium supernatant, etc. The purified RNA can be used for clinical in vitro detection.

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 Column Binding Capacity	Up to 50 µg RNA
Maximum Loading Volume Per Spin Column	650 µL
Size of RNA Purified	All sizes, including small RNA (< 200 nt)
Time to Complete 10 Purifications	1-4 hours*
Maximum Amount of Starting Material	5 slices of < 20 µm thick paraffin slices 25 mg of unsectioned block
Average Yield	Variable due to age of paraffin blocks ~2-3 µg of Total RNA per 1 mg of fresh FFPE hamster kidney

* Time required for purification varies by length of Proteinase K incubation and formalin crosslink-reversal

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