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).

Introduction

This product is designed for purification of high-molecular-weight genomic or mitochondrial DNA from a variety of blood samples. High-quality DNA can be purified from sample types including whole blood, buffy coat, bone marrow, body fluids in as little as 25 minutes. The convenient, scalable purification procedure removes contaminants and enzyme inhibitors such as proteins and divalent cation, and purified DNA is ready for immediate use in sensitive downstream applications or for archiving.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from whole blood using economic salt out method
Applications	PCR, enzyme digestion, Southern hybridization,etc.
Purification technology	Salting out method
Process method	Manual (centrifugation or vacuum)
Sample type	Animal tissue
Sample amount	Unlimited
Elution volume	≥100μl
Time per run	Variation with sample amount

Principles

Cells are lysed with ananionic detergent in the presence of a DNA stabilizer. The DNA stabilizer limits the activity of intracellular DNases and also DNases found elsewhere in the environment. RNA is then removed by treatment with an RNA digesting enzyme. Other contaminants, such as proteins, are removed by salt precipitation. Finally, the genomic DNA is recovered by precipitation with alcohol and dissolved in Buffer TE. Purified DNA typically has an A260/A280 ratio between 1.7 and 1.9, and is up to 200 kb in size. The DNA can be safely stored at 2-8°C, -20°C, or -80°C.

Advantages

• Safe – without phenol chloroform extraction
• Environment friendly – the reagents used are safe, non-toxic and without pollution
• High molecular weight – the molecular weight of genomic DNA is about 50-150kb
• High purity – the purified DNA has A260/280=1.8-1.9, A260/230=2.0-2.5
• Unlimited sample size – solution type operation, sample volume can be adjusted at will
• Cost performance – the most economical nucleic acid extraction technology

Kit Contents

Contents	D331101	D331102	D331103
Purification Volumes	50 ml	300 ml	1000 ml
10 x Buffer RBC	20 ml	100 ml	3 x 100 ml
Buffer WTL	60 ml	350 ml	1000 ml
Buffer PPS	20 ml	120 ml	350 ml
RNase Solution	300 µl	1.8 ml	6 ml
Buffer TE	10 ml	30 ml	120 ml

Storage and Stability

RNase Solution should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. The entire kit can be stored at 2-8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.

This product is designed for purification of high-molecular-weight genomic or mitochondrial DNA from a variety of blood samples. High-quality DNA can be purified from sample types including whole blood, buffy coat, bone marrow, body fluids in as little as 25 minutes. The convenient, scalable purification procedure removes contaminants and enzyme inhibitors such as proteins and divalent cation, and purified DNA is ready for immediate use in sensitive downstream applications or for archiving.

Introduction

Free-circulating nucleic acids, such as tumor-specific extracellular DNA fragments and mRNAs in the blood or fetal nucleic acids in maternal blood, are present in serum or plasma usually as short fragments, <1000bp (DNA). HiPure Circulating DNA Mini Kit enables efficient purification of these circulating nucleic acids from human plasma, serum, or urine. Samples can be fresh or frozen (provided that they have not been frozen and thawed more than once).

Details

Specifications

Features	Specifications
Main Functions	Isolation circulating DNA from 1ml plasma, serum, body fluids
Applications	qPCR, liquid or solid chip analysis, hybridization and SNP detection, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Serum, plasma and other cell-free fluid samples
Sample amount	1ml
Elution volume	≥30μl
Time per run	≤50 minutes
Liquid carrying volume per column	800µl
Binding yield of column	100µg

Principle

This product is based on silica gel purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption plate and filter column. Nucleic acid is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, nucleic acid was finally eluted with low-salt buffer (10 Mm Tris,pH 8.0).

Advantages

• High yield – most optimal process, free DNA (>50bp) can be obtained to the maximum extent
• High concentration – low elution volume, ensuring high nucleic acid concentration
• High purity – low alcohol binding method, completely removing inhibitor and protein pollution
• High recovery – DNA can be recovered at the level of PG by silica gel column purification

Kit Contents

Contents	D318102	D318103
Purification Times	50 Preps	250 Preps
Buffer ACL	50 ml	250 ml
Buffer ACB*	60 ml	300 ml
Buffer DCW1*	22 ml	88 ml
Buffer DCW2*	10 ml	50 ml
Proteinase K	120 mg	540 mg
Protease Dissolve Buffer	10 ml	30 ml
Carrier RNA	110 μg	310 μg
Nuclease Free Water	10 ml	30 ml
HiPure CFDNA Mini Columns	50	250
2 ml Collection Tubes	100	500

Storage and Stability

Proteinase K and 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 remainingkit components can be stored dry at room temperature (15–25°C) and are stable for at least 18 months under these conditions.The entire kit can be stored at 2–8°C, but in this case buffers shouldbe redissolved before use. Make sure that all buffers are at room temperature when used.

Free-circulating nucleic acids, such as tumor-specific extracellular DNA fragments and mRNAs in the blood or fetal nucleic acids in maternal blood, are present in serum or plasma usually as short fragments,