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 whole blood (fresh or frozen), plasma, serum, buffy coat, bone marrow, other body fluids, lymphocytes, cultured cells.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from 200ul Whole Blood
Applications	PCR, southern bolt and virus detection, etc
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Whole Blood (fresh or frozen), serum, plasma, milk, saliva, and other liquid samples and cultured cells
Sample amount	<200μl whole blood or other liquid samples, <5*106 lymphocytes or Culture CellsNon mammalian animals that have nucleus in red blood cells (rich in DNA, such as birds and fish) : 5~20μl whole blood at a time
Elution volume	≥20μl
Time per run	≤30 minutes
Liquid carrying volume per column	800µl
Binding yield of column	100µg

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.
• 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	D311102	D311103
Purification Times	50	250
HiPure DNA Mini Columns I	50	2 x 125
2ml Collection Tubes	100	5 x 100
Buffer AL	15 ml	60 ml
Buffer DW1	30 ml	150 ml
Buffer GW2*	12 ml	50 ml
Proteinase K	24 mg	120 mg
Protease Dissolve Buffer	1.8 ml	10 ml
Buffer AE	15 ml	60 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:

Introduction

Intended Use

For the detection of Salmonella spp. in food, animal feed and in environmental samples from the food production area as described in ISO 6579-1:2017.

Principle and Interpretation

Meat extract and casein provide a source of nitrogen and amino acids and sodium chloride maintain theosmotic balance. Ox bile and brilliant green act as selective agents against non-target microorganisms. Tetrathionate is generated from the sodium thiosulfate. Iodine and calcium carbonate buffer the sulfuric acid generated from tetrathionate reduction.

Formulation

Ingredients	/liter
Meat extract	4.3g
Enzymatic digest of casein	8.6g
Sodium chloride	2.6g
Calcium carbonate	38.7g
Sodium Thiosulfate (anhydrous)	30.4g
Ox bile	4.78g
pH8.0±0.2 at 25°C

Preparation

Suspend 89.4g in 1 L of purified water. Heat with frequent agitation and boil to completely dissolve the powder. Distribute into flasks，and then cool to below 45°C. Add a vial of novobiocin sodium salt (SR0640), a vial of iodine solution and a vial of brilliant green (SR0040) into 100 mL of base medium. Mix thoroughly.

Quality Control

Cultural characteristics observed after incubation at 35-37°C for 24 hours

Quality control strains	Approx. Inoculum(CFU)	Expected Results
Salmonella typhimurium  ATCC14028	10 – 100	≥ 10 cfu on XLD
Escherichia coli  ATCC25922	> 104	≤100 cfu on TSA
Enterococcus faecalis ATCC29212	> 104	<10 cfu on TSA

Storage and Shelf Life

2-30℃，Keep container tightly closed, avoid direct sunlight.

Use before expiry date on the label.

 Precautions

1. When weighing the dehydrated medium, please wear masks to avoid causing respiratory system discomfort

2. Keep container tightly closed after using to prevent clumping.

Waste Disposal

Microbiological contamination was disposed by autoclaving at 121°C for 30 minutes.

Revision

On June 14, 2024

References

ISO 6579:2017 Microbiology of food and animal feeding stuffs – Horizontal method for the detection, enumeration and serotyping of Salmonella spp.

Intended Use For the detection of Salmonella spp. in food, animal feed and in environmental samples from the food production area as described in ISO 6579-1:2017. Principle and Interpretation……

Hydroxy-Amino-bis(PEG2-propargyl) is a 3-arm linker with a terminal hydroxy group and two propargyl groups. The propargyl groups can participate in copper catalyzed azide-alkyne Click Chemistry to yield a stable triazole linkage. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Hydroxy-Amino-bis(PEG2-propargyl) is a 3-arm linker with a terminal hydroxy group and two propargyl groups. The propargyl groups can participate in copper catalyzed azide-alkyne Click Chemistry to yield a stable triazole linkage. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.