Introduction

Blood samples contain rich DNA, including mitochondrial DNA, genomic DNA, circulating DNA (mostly released into blood after tumor cell apoptosis) in white blood cells, as well as parasitic viral or microbial DNA. These DNA are important parameters in clinical testing or diagnosis, which are also valuable materials for medical research. There are three main issues with extracting DNA from blood samples:

1. The sample is highly infectious, posing great harm to operators and the environment.

2. The source of DNA is complex and aportion of the nucleic acid, such as viral DNA or free DNA, may be lost during the operation, leading to downstream detection failure;

3. Blood sample contains a large amount of impurities and inhibitory factors.

Currently there are many methods available for extracting DNA from whole blood samples, such as phenol chloroform extraction, salting out method, etc. However, these methods require pre-treatment of blood sample, which removes red blood cells and isolate white blood cells in the first step. Due to the requirement that it cannot inactivate or kill pathogens during the process of removing red blood cells, the waste liquid (red blood cell lysate) and consumables may be contaminated by pathogens and become infectious, posing a danger to the entire laboratory environment and operators. In addition, during the process of removing red blood cells, useful nucleic acid information such as viruses, microorganisms, or circulating DNA is also lost, leading to experiment or detection failures.

The HiPure Blood DNA Kits series provided by Magen Company uses silica gel column purification technology, which can directly lyse whole blood samples without the need for white blood cell separation. Whole blood samples are directly mixed with lysates and proteases, resulting in the inactivation of pathogens, greatly reducing the infectivity, environmental pollution, and the chance of operators being infected. Due to the direct lysis and digestion of samples, except lymphocyte DNA, other circulating DNA as well as DNA from viruses and microorganisms, can also be recovered.

This product provides fast and easy methods for purification of total DNA for reliable PCR and Southern Blotting. Total DNA (e.g., genomic, viral, mitochondrial) can be purified from tissue and culture cells.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from 2ml blood and 200mg tissue using Midi column
Applications	PCR, southern bolt and virus detection, etc
Purification method	Midi spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Tissue, cell, blood, saliva, swab, blood spot, semen and other clinical samples
Sample amount	0.2-2 ml
Elution volume	≥300μl
Time per run	≤80 minutes
Liquid carrying volume per column	4ml
Binding yield of column	1mg

Principles

This product is based on silica column purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption 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 (10mm Tris, pH9.0, 0.5mm EDTA).

Advantages

• High quality DNA – meet a variety of downstream applications, including PCR, qPCR, enzyme digestion, hybridization, etc.
• Fast – without separation of leukocytes, organic extraction or ethanol precipitation
• Simple – all nucleic acids can be obtained by direct digestion
• Wide applicability – handle a variety of liquid samples

Kit Contents

Contents	D311302	D311303
Purification Times	20	100
HiPure gDNA Midi Columns	20	100
15ml Collection Tubes	40	200
Buffer ATL	50 ml	250 ml
Buffer AL	50 ml	250 ml
Buffer GW1*	22 ml	110 ml
Buffer GW2*	12 ml	50 ml
RNase A	20 mg	90 mg
Proteinase K	100 mg	440 mg
Protease Dissolve Buffer	10 ml	30 ml
Buffer AE	20 ml	120 ml

Storage and Stability

Proteinase K, RNase A 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 conditions.

Purchase Guide

Name	CAT NO	Sample amount	Leukocyte protocol*	Colum type	Elutio volume	Average yield	Time per run
HiPure Blood DNA Mini Kit	D3111	10-200μl	1ml	2ml column	≥20μl	5-9μg/200μl	≤30 minutes
HiPure Tissue&Blood DNA Midi Kit	D3113	0.2-2ml	10ml	1.5ml column	≥300μl	20-40μg/1m	≤80 minutes
HiPure Tissue&Blood DNA Maxi Kit	D3115	3 -10ml	10ml	15ml column	≥700μl	20-40μg/1m	≤90 minutes
HiPure Tissue&Blood DNA 96 Kit	D3117	1-200μl	1ml	96 well plate		3-8μg/200μl

Blood samples contain rich DNA, including mitochondrial DNA, genomic DNA, circulating DNA (mostly released into blood after tumor cell apoptosis) in white blood cells, as well as parasitic viral or microbial DNA. These DNA are important parameters in clinical testing or diagnosis, which are also valuable materials for medical research. There are three main issues with extracting DNA from blood samples:

Overview

• Isolate all sizes of circulating and exosomal RNA, including microRNA
• Versatile plasma/serum input ranges
• No phenol extractions
• No carrier RNA
• Bind and elute all RNA irrespective of size or GC content, without bias
• Concentrate circulating RNA and exosomal RNA into a flexible elution volume
• High quality, purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services
• Compatible with Streck Cell-Free RNA BCT® Tubes
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

These kits provide a fast, reliable and convenient method to purify and concentrate high quality, high purity and inhibitor-free cell-free circulating and exosomal RNA using a convenient spin column method. These kits can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used, as heparin can significantly interfere with many downstream applications such as RT-PCR. The purified plasma/serum RNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive reverse transcription qPCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, expression array assays, and NGS.

Background

Plasma/Serum cell-free circulating RNA or exosomal RNA has the potential to provide biomarkers for certain cancers and disease states. Exosomes are 40 – 150 nm membrane vesicles, which are secreted by most cell types. Exosomes can be found in saliva, blood, urine, amniotic fluid and malignant ascitic fluids, among other biological fluids. Evidence has been accumulating recently that these vesicles act as cellular messengers, conveying information to distant cells and tissues within the body. These exosomes may play a functional role in mediating adaptive immune responses to infectious agents and tumours, tissue repair, neural communication and transfer of pathogenic proteins. For this reason exosomal RNAs may serve as biomarkers for various diseases including cancer. As the RNA molecules encapsulated within exosomes are protected from degradation by RNAses they can be efficiently recovered from biological fluids, such as plasma or serum.

Plasma/Serum RNA Purification Mini Kit

This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 50 µL to 200 µL. The purified plasma/serum RNA is eluted in a flexible final volume of 10 µL to 25 µL.

Plasma/Serum RNA Purification Midi Kit

This utilizes a two column method, and can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 250 µL to 1.5 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 µL to 100 µL.

Plasma/Serum RNA Purification Maxi Kit

This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 2 mL to 5 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 µL to 100 µL.

Isolate RNA after Purifying EVs and Exosomes

For Ultracentrifugation, Exoquick, and Filtration

Cat. #	Name	Elution Volume	Plasma/Serum	Urine	Cell-Culture Media
55000	Plasma/Serum RNA Purification Mini Kit	10 – 25 µL	50 µL – 1 mL	250 µL – 1 mL	5 – 10 mL
35300	Total RNA Purification Micro Kit	20 – 50 µL	1 – 4 mL	2 – 10 mL	10 – 20 mL
17200	Total RNA Purification Kit	50 – 100 µL	4 – 10 mL	11 – 30 mL	20 – 35 mL

Details

Supporting Data

Figure 1 / 13

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Kit Specifications
Sample Volume Range	50 to 200 μL
Anti-coagulant (for Plasma)*	EDTA or Citrate
Size of RNA Purified	All sizes, including small RNA (< 200 nt)
Minimum Elution Volume	10 μL
Maximum Elution Volume	25 μL
Time to Complete 10 Purifications	15-20 minutes
Average Yield**	Variable depending on specimen

*This kit is suitable for the isolation of RNA from fresh or frozen serum or plasma prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used as heparin can significantly interfere with many downstream applications such as RT-PCR.

**Please check page 7 for Average Plasma/Serum Yields and Common RNA Quantification Methods.

Storage Conditions and Product Stability

All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.

Kit Components	Cat. 55000 (50 preps)	Cat. 56100 (20 preps)	Cat. 56200 (10 preps)
Lysis Buffer A	2 x 20 mL	100 mL	1 x 130 mL 1 x 30 mL
Wash Solution A	18 mL	1 x 38 mL 1 x 18 mL	38 mL
Elution Solution A	6 mL	6 mL	6 mL
Elution Buffer F	–	15 mL	15 mL
Micro Spin Columns	50	–	–
Mini Spin Columns	–	20	10
Midi Spin Columns	–	20	–
Maxi Spin Columns	–	–	10
Collection Tubes	50	20	10
Elution Tubes (1.7 mL)	50	20	10
Product Insert	1	1	1

Overview

• For use from preserved saliva collected with Norgen’s Saliva RNA Collection and Preservation Devices Dx (Cat. 53800) or preserved swabs collected in Norgen’s Total Nucleic Acid Preservative Tubes Dx (Cat. Dx69200)
• CE-IVDR marked in accordance with the European Commission Regulation (EU) No. 2017/746.
• Fits into in vitro diagnostic workflows
• Isolate high-quality total RNA, including viral RNA, from fresh and preserved saliva and swab samples
• Fast and easy processing using rapid spin column format
• Isolate total RNA, from large rRNA down to microRNA (miRNA)
• No phenol or chloroform extractions
• Very sensitive & linear down to a few cells/viral copies without the need for carrier RNA
• Buffer chemistry inactivates viruses including SARS-CoV-2 – Explore the Application Note
• 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

These kits provides a rapid method for the purification of total RNA from non-preserved saliva and nasal/throat swabs, and from preserved saliva collected on Norgen’s Saliva RNA Collection and Preservation Devices Dx (Cat. 53800) or preserved swabs collected in Norgen’s Total Nucleic Acid Preservative Tubes Dx (Cat. Dx69200). Purification is based on using Norgen’s proprietary resin separation matrix. RNA is preferentially purified from other cellular components such as proteins, without the use of phenol or chloroform. The chemistry employed in the kit allows the purification of total RNA, including viral and bacterial RNA, irrespective of size or GC content. The purified RNA is ideal for in vitro diagnostic use for medical purposes.

Saliva/Swab RNA Purification Kit Dx (Spin Column)

This kit is optimized to be used with any downstream application employing enzymatic amplification or other enzymatic modifications of RNA followed by signal detection or amplification. Any diagnostic results generated using the RNA isolated with Norgen’s Saliva / Swab RNA Purification Kit Dx in conjunction with an in vitro diagnostic assay should be interpreted with regard to other clinical or laboratory findings.

To minimize irregularities in diagnostic results, suitable controls for downstream applications should be used.

Norgen’s Saliva/Swab RNA Purification Kit Dx is intended for use by professional users such as technicians, physicians and biologists experienced and trained in molecular biological techniques.

Norgen’s Saliva/Swab RNA Purification Kit Dx does not provide a diagnostic result. It is the sole responsibility of the user to use and validate the kit in conjunction with a downstream in vitro diagnostic assay.

Saliva/Swab RNA Purification 96-Well Kit Dx (High Throughput)

This kit is optimized to be used with any downstream application employing enzymatic amplification or other enzymatic modifications of RNA followed by signal detection or amplification. Any diagnostic results generated using the RNA isolated with Norgen’s Saliva / Swab RNA Purification 96-Well Kit Dx in conjunction with an in vitro diagnostic assay should be interpreted with regard to other clinical or laboratory findings. To minimize irregularities in diagnostic results, suitable controls for downstream applications should be used. Norgen’s Saliva / Swab RNA Purification 96-Well Kit Dx is intended for use by professional users such as technicians, physicians and biologists experienced and trained in molecular biological techniques. Norgen’s Saliva / Swab RNA Purification 96-Well Kit Dx does not provide a diagnostic result. It is the sole responsibility of the user to use and validate the kit in conjunction with a downstream in vitro diagnostic assay.

• High throughput 96-well plate format for rapid purifications
• Very consistent well-to-well RNA isolation
• Bind & elute all RNA irrespective of size or GC content, without bias

Details

Supporting Data

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Kit Specifications
Sample Volume Range	250 µL
Size of RNA purified	All sizes, including small RNA (<200 nt)
Minimum Elution Volume	75 µL
Maximum Elution Volume	100 µL
Time to complete 96 purifications	30 minutes
Average Yield	≥ 1 µg **varies from sample to sample

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature.  All solutions and plastics can be used until the expiration date specified on their labels. 

Component	Cat. Dx69100 (50 preps)	Cat. Dx69300 (192 preps)
Lysis Buffer A	30 mL	100 mL
Solution WN	18 mL	55 mL
Wash Solution A	18 mL	2 x 38 mL
Elution Solution A	6 mL	20 mL
Mini Spin Columns	50	–
96-Well Isolation Plate (Deep Well)	–	2
Collection Tubes	50	–
96-Well Collection Plate (Deep Well)	–	2
Elution Tubes (1.7 mL)	50	–
96-Well Elution Plate (Deep Well)	–	2
Adhesive Tape	–	4
Product Insert	1	1