Overview

• Simple protocol that is free of detergents
• Fast processing time
• Process a variety of inputs
• Compatible with mass spectrometry and NMR spectroscopy
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

This kit provides a fast and simple procedure for the isolation of total proteins from tissue, bacteria, yeast or mammalian cells without the use of SDS, Triton® X-100 and other detergents. Detergents are extensively used to prepare protein samples; however, these detergents have undesirable effects on downstream analysis. These effects include extraneous peaks in mass spectrometry, artifacts with chromatography and electrophoresis, interference with microinjection into cells and interference with protein immunization. 

The Detergent-Free Total Protein Isolation Kit maintains high protein recovery and yields proteins that are 100% detergent-free.  Purification is based on using Norgen’s proprietary resin together with Lysis Solution, followed by protein filtration using a filter column (provided). The purified proteins can be used in a number of downstream applications including mass spectrometry, SDS-PAGE, isoelectric focusing, NMR spectroscopy and more.

Each Lysis Tube is able to process up to 50 mg of tissue, 10¹⁰ bacterial cells, 10⁹ yeast cells or 10⁷ mammalian cells. Preparation time for 12 samples is less than 10 minutes. 

Details

Supporting Data

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Kit Specifications
Maximum Amount Of Starting Material: Tissues Animal Cells Bacteria Yeast	50 mg 1 x 107 cells 1 x 1010 cells 1 x 109 cells
Time to Process 12 Samples	Less than 10 minutes

Storage Conditions and Product Stability
The Lysis Solution should be kept tightly sealed and stored at room temperature. Once opened, the solution should be stored at 4°C. This kit is stable for 2 years after the date of shipment.

Component	Cat. 30300 (50 preps)
Lysis Solution	4 mL
Lysis Tubes	25
Filter Columns	25
Elution Tubes (1.7 mL)	25
Product Insert	1

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 provide fast and easy methods for purification of total DNA from whole blood, plasma, serum, buffy coat, bone marrow, other body fluids, lymphocytes, cultured cells. There is no need to use toxic phenol chloroform extraction or time-consuming alcohol precipitation. The extraction process finish in 40 minutes. Purified DNA includes genomic DNA, mitochondrial DNA, viral DNA (e.g. HBV), or DNA from other parasitic microorganisms. The obtained DNA can be directly used in PCR, viral DNA detection and other experiments.

This kit can use on manual protocol or 16/32/48 channel automated extraction system. No need to pause for the addition of binding solution (not applicable to 96/192 instruments)

Details

Kit Contents

Contents	D631001C	D631002C	D631003C
Purification Times	48	96	480
MagPure Particles	1.7 ml	3.5 ml	16 ml
Proteinase K	24 mg	50 mg	220 mg
Protease Dissolve Buffer	1.8 ml	3 ml	15 ml
Buffer MLA	30 ml	70 ml	300 ml
Buffer DW1	30 ml	70 ml	300 ml
Buffer EW	60 ml	120 ml	2 x 300 ml
Buffer MWX1	30 ml	70 ml	300 ml
Elution Buffer	10 ml	20 ml	60 ml

Prefilled Kit Contents

Product	Contents and volume	D6310-TL-06C	D6310-S-48
Proteinase K		50 mg	24 mg
Protease Dissolve Buffer		3 ml	1.8 ml
TL-Tip		12 pcs	24 pcs
Tip bottom plate/ Tip base reagent strip	Row 1/7：600μl Buffer MLA	6 plates	48 strips
	Row 2/8：600μl Buffer MWX1
	Row 3/9：600μl Buffer DW1
	Row 4/10：30μl MagPure Particles 600μl Buffer EW
	Row 5/11：600μl Buffer EW
	Row 6/12：100μl Elution Buffer

Storage and Stability

Proteinase K Powder 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 these condition.

This product provide fast and easy methods for purification of total DNA from whole blood, plasma, serum, buffy coat, bone marrow, other body fluids, lymphocytes, cultured cells. There is no need to use toxic phenol chloroform extraction or time-consuming alcohol precipitation. The extraction process finish in 40 minutes. Purified DNA includes genomic DNA, mitochondrial DNA, viral DNA (e.g. HBV), or DNA from other parasitic microorganisms. The obtained DNA can be directly used in PCR, viral DNA detection and other experiments.