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

• Fractionate leukocytes from whole blood in minutes with provided RBC Lysis Buffer
• Isolate total RNA, including microRNA, without phenol
• Rapid and convenient spin-column format and 96-well plate for high throughput applications
• Purified RNA is ready for any downstream application including RT-PCR, qRT-PCR, NGS, arrays and more. 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 provide a rapid method for the isolation and purification of total leukocyte (white blood cell) RNA from mammalian blood samples. These kits are supplied with an RBC (red blood cell) Lysis Buffer for selective removal of red blood cells and fractionation of leukocytes by centrifugation. Isolation of leukocyte RNA results in improved expression profiling and other downstream applications by removing the masking effects of some RNAs which are very abundant in whole blood, such as globin mRNAs. These kits are able to isolate total leukocyte RNA, including both large mRNA and all small RNA species containing microRNA (miRNA) and small silencing RNA (siRNA). The purified RNA is of the highest quality and can be used in a number of downstream applications.

Leukocyte RNA Purification Kit (Spin Column)

This kit provides a rapid method for the isolation and purification of total leukocyte RNA from mammalian blood samples in 40 minutes. Allowable blood input ranges from 10 μL to 2 mL or 3 x 10⁶ Leukocytes

Leukocyte RNA Purification Plus Kit (Plus)

Norgen’s Leukocyte RNA Purification Plus Kit provides a rapid method for the isolation and purification of total leukocyte (white blood cell) RNA from up to 3 mL of mammalian blood samples. Complete 10 purifications in as little as 40 minutes.

Leukocyte 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. Norgen’s kit allows for the isolation of total leukocyte RNA, including all small RNA species. The purified RNA is of the highest quality and can be used in a number of downstream applications including real time PCR, reverse transcription PCR, northern blotting, RNase protection and primer extension, and expression array assays. Allowable blood input ranges from 10 μL to 1 mL or 1.5 x 10⁶ Leukocytes.

Details

Supporting Data

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Kit Specifications
Maximum Binding Capacity Per Well	50 μg
Maximum Loading Volume Per Well	500 μL
Size of RNA Purified	All sizes, including small RNA < 200 nt
Maximum Blood Input*	1 mL or 1.5 x 106 Leukocytes
Standard Blood Input	150 μL
Minimum Blood Input	10 μL
Average Yield: 500 μL human blood	1.5 μg
Time to Complete 10 Purification	40 minutes

*Additional RBC Lysis Solution is required for input Volumes >150 µL

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature.  The RBC Lysis Buffer should be stored at 4°C upon arrival.  These reagents should remain stable for at least 1 year in their unopened containers.

Component	Cat. 21200 (50 preps)	Cat. 21250 (50 preps)	Cat. 37800 (2 x 96 preps)
RBC Lysis Buffer	2 x 100 mL	2 x 1 L	2 x 90 mL
Buffer RL	30 mL	30 mL	–
Binding Solution	–	–	2 x 40 mL
Wash Solution A	38 mL	38 mL	–
Wash Solution	–	–	2 x 30 mL
Elution Solution A	6 mL	6 mL	–
Elution Solution	–	–	2 x 20 mL
Enzyme Incubation Buffer	–	6 mL	1
DNase I	–	1 Vial	–
Mini Spin Columns	50	–	–
Lysate Homogenization Column	–	50	–
Single Cell RNA Column	–	50	–
RBC Lysis 96-Well Plate	–	–	2
96-Well Filter Plate	–	–	2
Adhesive Tape	–	–	4
Collection Tubes	50	100	–
96-Well Collection Plate	–	–	2
Elution Tubes (1.7 mL)	50	50	–
96-Well Elution Plate	–	–	2
Product Insert	1	1	1