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, whole blood, plasma, serum, buffy coat, bone marrow, other body fluids, lymphocytes, cultured cells.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from blood, tissue, culture cells, swab, blood spots using 96 plate
Applications	PCR, southern bolt and virus detection, etc
Purification method	96 well plate
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Blood, serum, plasma, milk, saliva, and other liquid samples and cultured cells
Sample amount
Elution volume
Time per run
Liquid carrying volume per column
Binding yield of column

Principle

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.
• High throughput – 96 samples can be processed simultaneously

Kit Contents

Contents	D311701	D311702
Purification Times	1 x 96	4 x 96
HiPure gDNA Plate	1	4
96 well Plate (2.2ml)	1	4
1.6ml Collection Plate	1	4
0.5ml Collection Plate	1	4
Silicon Seal Tape	1	4
Seal Film	5	25
Buffer ATL	30 ml	100 ml
Buffer AL	30 ml	100 ml
Buffer DW1	60 ml	250 ml
Buffer GW2	50 ml	2 x 100 ml
Proteinase K	50 ml	200 ml
Protease Dissolve Buffer	5 ml	15 ml
Buffer AE	30 ml	120 ml

Storage and Stability

Proteinase K 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:

N-(Amino-PEG4)-N-bis(PEG4-propargyl) HCl salt is a trifunctional linker with two terminal alkynes and a primary amine. The terminal alkynes can be linked to azides using copper click chemistry while the primary amine is reactive towards carboxylic acids, NHS esters, ketones and aldehydes to form a variety of stable bonds.

N-(Amino-PEG4)-N-bis(PEG4-propargyl) HCl salt is a trifunctional linker with two terminal alkynes and a primary amine. The terminal alkynes can be linked to azides using copper click chemistry while the primary amine is reactive towards carboxylic acids, NHS esters, ketones and aldehydes to form a variety of stable bonds.

Overview

• Rapid AAV DNA extraction and quantification can be completed in 2 hours 
• Universal qPCR assay targeting the AAV2 ITR enables quantification of most AAV vector constructs
• Purified DNA standard enables easy comparison of data between different lab groups
• Non-plasmid based standard prevents quantification artifacts due to insufficient melting of plasmid sequences
• DNAse digestion step aids in eliminating non-viral DNA
• Minimal sample handling for maximum AAV DNA recovery

Norgen’s AAV Quantification Kit is designed for the detection of adeno-associated virus (AAV) inverted terminal repeat (ITR) sequences in a real-time PCR based on the use of TaqMan® technology. A purified standard based solely on the AAV2 ITR sequence purified using Norgen’s proprietary silicon carbide technology simplifies the generation of a reliable standard curve for AAV quantification. Avoidance of a plasmid based standard eliminates problems associated with efficient melting of the ITR sequence due to coupling of the ITR to the much longer plasmid sequence, as well as variability due to rearrangements/duplications/deletions of the recombination prone ITR. An easy and rapid method for viral DNA extraction simplifies the step of obtaining AAV DNA while simultaneously eliminating contaminating non-encapsidated DNA.  Norgen’s AAV Quantification Kit can facilitate pre-clinical studies that require accurate vector titration as well as interlab comparisons of vector quantities. This kit is designed for research use only and not for use in diagnostic procedures.

Details

Supporting Data

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Storage Conditions and Product Stability
All kit components should be stored at -20°C upon arrival. Repeated thawing and freezing (> 2 x) of the Master Mix and Positive Control should be avoided, as this may affect the performance of the assay. If the reagents are to be used only intermittently, they should be frozen in aliquots. All reagents can be stored for 1 year at -20°C without showing any reduction in performance.

Component	Cat. 63800 (24 rxns)
Enzyme Incubation Buffer A	500 µL
DNase I	25 µL
DNase Stop Solution	100 µL
2X PCR Mastermix	350 µL
AAV Primer & Probe Mix	90 µL
AAV Standard	6 µL
Nuclease-Free Water (Negative Control)	1.25 mL
Product Insert	1