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

• Preservation and isolation of both cf-DNA and cf-RNA from a single tube
• Fixative-free preservative, no cross-linking of DNA
• Preserve cf-DNA/ct-DNA for 30 days at ambient temperature and for up to 8 days at 37°C
• Preserve cf-RNA for 30 days at ambient temperature
• Preserve Circulating Tumour Cells (CTCs) for 14 days at ambient temperature
• No plasma volume loss after shipping/transportation
• Prevent hemolysis allowing better separation of plasma
• Prevent apoptosis of blood cells and fragmentation of genomic DNA
• Produce high quality/quantity of plasma cf-DNA/ct-DNA/cf-RNA
• Vacuumed to draw 8.7 mL of blood in 10 mL tubes
• The procedure for plasma isolation using this kit is compliant with ISO 20186-3:2019

Norgen’s cf-DNA/cf-RNA Preservative Tubes are closed, evacuated plastic tubes for the collection and the preservation of cf-DNA, circulating tumor DNA, cf-RNA and circulating tumor cells in human whole blood samples during storage and shipping. Using these tubes in combination with Norgen’s Plasma/Serum Cell-Free Circulating DNA Purification Kits or Norgen’s Plasma/Serum RNA Purification Kits enables the purification and concentration of an inhibitor-free cf-DNA/ct-DNA/cf-RNA in a very small elution volume. Plasma recovered from Norgen’s cf-DNA/cf-RNA Preservative Tubes is also compatible with any other cf-DNA and/or cf-RNA purification method. The purified cf-DNA, ct-DNA and cf-RNA are compatible with any downstream application assays, including PCR, qPCR, rt-qPCR, methylation-sensitive PCR, Southern Blot analysis, gene expression analysis, microarrays and NGS.

This product is for research use only and not for use in diagnostic procedures.

Performance

Norgen’s cf-DNA/cf-RNA Preservative Tubes abridge the collection/processing of whole blood for the subsequent purification of cf-DNA and/or cf-RNA. The cf-DNA/cf-RNA Preservative Tubes preserve and inhibit the programmed cell-death (apoptosis) of the blood, hence preventing the release of intracellular DNA/RNA into plasma. Whole blood samples can be shipped and stored at room temperature for the subsequent processing of plasma. The cf/ct DNA and cf-RNA levels are stable for up to 30 days at room temperature. The cf-DNA is also stable for up to 8 days at 37°C. When blood collected on Norgen’s cf-DNA/cf-RNA tubes is processed for plasma recovery no buffy coat will be generated and the Circulating tumor cells (CTCs) will be located at the bottom of the tube with the blood sediment. CTCs are stable for 14 days at room temperature where it can be extracted from the blood sediment or from whole preserved blood for cell sorting or for cf/DNA or cf-RNA purification.

Plasma samples produced from the whole blood sample collected into Norgen’s cf-DNA/cf-RNA Preservative Tubes and purified using Norgen’s Plasma/Serum Cell-Free Circulating DNA Purification Kits or Norgen’s Plasma/Serum RNA Purification Kits yield concentrated, high-quality and inhibitor-free cf-DNA/ct-DNA or cf-RNA for any downstream application.

Workflow

NOTE: This product is for research use only. For the CE marked version, see Cat. Dx63950

Details

Supporting Data

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Kit Specifications
Tube Size	16 x 100 mm
Tube Type	PET Tube
Blood Draw Volume	8.7 mL in 10 mL Tubes
Preservative Volume	1.3 mL
Recovered Plasma Volume	Approximately 6-7 mL (not affected by shipping)
Anticoagulant	Proprietary
Preservative	Non formaldehyde – Proprietary preservation agent
Length of cf-DNA Preservation	Up to 30 days at room temperature (15-25°C) Up to 8 days at 37°C
Length of cf-RNA Preservation	Up to 30 days at room temperature (15-25°C)
Length of CTCs Preservation	Up to 14 days at room temperature (15-25°C)

Storage Conditions and Product Stability

1. When stored at 15-30°C, unfilled Norgen cf-DNA/cf-RNA Preservative Tubes are stable through the expiration date.  See tube label for expiry date.
2. Short-term storage/shipping at -20°C to 37°C for up to 10 days is acceptable for unfilled Norgen cf-DNA/cf-RNA Preservative Tubes.
3. Do not freeze unfilled Norgen cf-DNA/cf-RNA Preservative Tubes for long term storage over 10 days. Freezing unfilled tubes may lead to loss of vacuum and precipitation may occur.
4. Cell-free DNA in blood samples collected in Norgen’s cf-DNA/cf-RNA Preservative Tubes is stable for up to 30 days when stored at room temperature (15-25°C)
5. Cell-free DNA in blood samples collected in Norgen’s cf-DNA/cf-RNA Preservative Tubes is stable for up to 8 days when stored at 37ºC.
6. Cell-free RNA in blood samples collected in Norgen’s cf-DNA/cf-RNA Preservative Tubes is stable for up to 30 days when stored at room temperature (15-25°C)
7. Circulating tumor cells (CTCs) in blood samples collected in Norgen’s cf-DNA/cf-RNA Preservative Tubes are stable for up to 14 days at room temperature (15-25ºC).

Product Description

Product Detail

Kit Storage and term of Validity

Storage term: stored at ≤-20℃,keep away from light, avoid heavy weight and repeated freezing and thawing.

Term of Validity: 14 months

Isothermal nucleic acid Principle Summary

This kit is based on a rapid nucleic acid amplification technology at room temperature and constant temperature: at room temperature and constant temperature (generally 39 ºC~42 ºC), reverse transcriptase uses specific primers and template RNA to synthesize cDNA strands, and binds the auxiliary protein and single strand With the help of the protein, the recombinase and the primer form a complex; perform a homology search and bind the target homology domain, at this time a D-loop region is formed at the homology position and strand exchange begins; accompanied by the recombinase from the complex Upon dissociation, the polymerase also binds to the 3′ end of the primer, initiating chain extension. Relying on the action of nfo enzyme, adding specific molecular probes designed according to the template, and using colloidal gold technology (sandwich method) can detect the final result.

Isothermal nucleic acid Product Features

1/ High sensitivity and specificity, short reaction time.

2/ The reagent form is freeze-dried, stable and easy to operate.

3/ The reaction can be operated by metal bath and water bath pot without purchasing expensive PCR apparatus.

Technical Parameters:

Parameters	Details
Product Name	RNA Isothermal Amplification Kit NFO
Manufacturer	Amp-future
Storage Temperature	-20°C
Kit Components	Enzymes, Buffers ,Reagents
Packaging	48 Tests/box
Detection Limit	500-1000copies/µL
Shipping	ICE
Test Time	5-20mins

Isothermal nucleic acid Applications

Suitable for RNA isothermal rapid amplification kit(NFO type)

Primer: Require pair of nucleotide primers with the length of 25-35 bp.

RNA NFO kit reaction temperature is 39 to 42℃ and time is 5-20 minutes.

Notes

1/ Please avoid nucleic acid contamination and set blank control during reaction due to the high sensitivity of the kit.

2/ Please take out the required quantity of MIRA reaction units for the experiment, and put the rest under storage conditions when performing the experiment.

Storage term: stored at ≤-20℃,keep away from light, avoid heavy weight and repeated freezing and thawing.

Term of Validity: 14 months