Overview

• Detection kits for Clostridium difficile
• CE-IVD marked version available for in vitro diagnostic use
• Available in TaqMan format for analysis

Clostridium difficile is rod-shaped, gram positive bacterium. It is the main causal agent of antibiotic-associated diarrhea and pseudomembranous colitis. The colonization of intestines by C. difficile is usually associated with the elimination of natural intestinal flora as a result of antibiotic application and is frequently reported in health care centers. While C. difficile infection could be severe and life-threatening, particularly among the elderly, many patients are asymptomatic making diagnosis challenging during outbreaks. The tradition method of detecting C. difficile infection is by cytotoxicity test of the toxin produced by the bacterium, but such protocols usually require extensive time before conclusion can be made.

Details

Supporting Data

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Storage Conditions and Product Stability
All kit components can be stored for 2 years after the date of production without showing any reduction in performance.

All kit components should be stored at -20°C upon arrival.

Component	Cat. TM37150 (100 preps)	Cat. TM37110 (100 preps)
MDx TaqMan 2X PCR Master Mix	2 x 700 μL	–
Clostridium difficile Primer & Probe Mix	280 μL	280 μL
Clostridium difficile Positive Control	150 μL	150 μL
Nuclease-Free Water (Negative Control)	1.25 mL	1.25 mL
Product Insert	1	1

Overview

• Efficient depletion of bovine exosomes from Fetal Bovine Serum
• Deplete exosome-sized vesicles from versatile FBS volumes
• No protease treatment required
• No time-consuming ultracentrifugation
• No precipitation reagents required
• No overnight incubation required
• Exosome depletion confirmed by reduction of bovine miRNAs below detectable levels
• The depleted FBS provides the same cellular growth rates as the standard FBS
• Purification is based on Norgen’s proprietary Silicon Carbide resin matrix

Norgen’s FBS Exosome Depletion Kits provides a quick and easy protocol for the depletion of bovine exosomes from FBS prior to using it as a growth supplement in your culture medium. The FBS recovered from the depletion process is exosome-depleted and does not contain any quantifiable bovine miRNAs. Moreover, the exosome-depleted FBS will support the growth of your cells of interest similar to the non-depleted FBS. Norgen’s kits allow for the processing of different FBS volumes. The depletion is based on Norgen’s proprietary resin. These kits provide a clear advantage over other available kits in that they do not require ultracentrifugation, any special instrumentation, precipitation reagents or any protease treatments. More importantly, the depletion process is an inexpensive method for exosome depletion from FBS, as compared to the expensive current ready-to-use exosome-depleted media available on the market.

Background

Most culture medium used for the growth and propagation of cells in culture require the addition of fetal bovine serum (FBS) as a growth supplement to media. FBS is obtained from bovine (cow) serum, and therefore contains large quantities of bovine exosome vesicles. These exosomes may interfere with some types of studies, or may lead to unreliable results when studying the exosomes shed from your cells of interest in normal culture conditions. Therefore, the use of exosome-depleted FBS is highly recommended for many types of studies.

FBS Exosome Depletion Kit (Slurry Format)

For FBS volumes ranging from 140 mL to 280 mL.

FBS Exosome Depletion Kit (Column Format)

For FBS volumes ranging from 120 mL to 240 mL.

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Supporting Data

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Kit Specifications
Sample Type	Fetal Bovine Serum
Sample Volume Range	Up to 140 mL (FBS Exosome Depletion Kit I (Slurry Format) Up to 280 mL (FBS Exosome Depletion Kit II (Slurry Format)
Depletion	Deplete exosome-sized vesicle
Bovine miRNA	No detectable bovine miRNA
Time to Complete 6 Purifications	40 minutes

Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.

Component	Cat. 61100 (6 preps)	Cat. 61400 (12 preps)	Cat. 61200 (6 preps)	Cat. 61300 (12 preps)
ExoC Buffer	2 x 1.5 mL	8 mL	2 x 1.5 mL	8 mL
Slurry E	12.5 mL	2 x 12.5 mL	–	–
Maxi Spin Column	–	–	6	12
Product Insert	1	1	1	1

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, tissue and culture cells.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from 10ml blood and 1g tissue using Maxi column
Applications	PCR, southern bolt and virus detection, etc
Purification method	Maxi 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	3-10ml
Elution volume	≥700μl
Time per run	≤90 minutes
Liquid carrying volume per column	4ml
Binding yield of column	5mg

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
• Pertinence – specially designed for isolating DNA from 3-10ml blood and related body fluids
• Wide applicability – handle a variety of liquid samples
• Economy – excellent cost effectiveness performance

Kit Contents

Contents	D311502	D311503
Purification Times	10	50
HiPure gDNA Maxi Columns	10	50
50ml Collection Tubes	20	100
Buffer ATL	120 ml	550 ml
Buffer AL	120 ml	2 x 300 ml
Buffer GW1*	53 ml	220 ml
Buffer GW2*	25 ml	110 ml
RNase A	40 mg	180 ml
Proteinase K	120 mg	540 mg
Protease Dissolve Buffer	12 ml	50 ml
Buffer AE	30 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: