Overview

• Use with Norgen’s TaqMan Probe/Primer and Control Sets
• For any PCR application where the input template quality is important to avoid false negatives
• Convenience and time savings
• Cost efficient
• High sensitivity

TaqMan 2x PCR Master Mix – Cat. 28340

Norgen’s TaqMan 2x PCR Master Mix is a ready-to-use TaqMan 2x PCR Master Mix solution that contains a PCR internal control which can be detected by HEX/VIC channel in a real-time PCR machine. By detecting the internal control users can validate the DNA template quality, thereby preventing any false negatives in the PCR results. The user needs only to add template, target TaqMan primer/probe mix and water to set up the TaqMan real-time PCR. 

PCR Control: 
TaqMan 2x PCR Master Mix contains PCR control primers/probe (HEX/VIC) and PCR control template. The PCR control reaction in the TaqMan 2x PCR Master Mix is optimized to not interfere with target amplification. The fluorescence of the target probe should not be HEX/VIC.

Details

Reagents Supplied – Cat # 28340

• TaqMan 2x PCR Master Mix (3 Vials, 100 Reactions) – Sufficient reagent for 100 x 20 µL reactions

Storage Conditions and Product Stability
Norgen’s TaqMan 2x PCR and 2x RT-PCR Master Mixes should be stored at -20ºC. For everyday use an aliquot can be stored at 4ºC for up to three months. The Master Mix is stable for multiple freeze-thaw cycles. When stored at the proper temperature this reagent is stable for at least 1 year.

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

*Note：Leukocyte protocol can be used when large volume whole blood samples need to be processed. Whole blood was treated with red blood cell lysate, and white blood cells were obtained by centrifugation before extraction

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

• Detection kits for the HSV-1
• CE-IVD marked version available for in vitro diagnostic use
• Available in TaqMan format for analysis

Herpes Simplex Virus 1 (HSV-1) is a member of the herpes virus family, Herpesviridae. HSV-1 has a relatively large double-stranded DNA genome. HSVs are primarily transmitted by sexual intercourse, direct contact with lesions or perinatally. Most HSV positive cases are characterised by lesions on the skins and mucous membranes of the mouth and genitals. HSV infection can be either primary or a recurrence of a previous infection. More than 90% of the primary HSV infections are asymptomatic. Primary infection with HSV-1 can lead to gingivostomatitis, eczema herpeticum, keratoconjunctivitis and encephalitis. The primary symptoms of a secondary infection are skin lesions in the nose, mouth and genital regions. The infection is contagious, mainly during an epidemic.

HSV-1 TaqMan PCR Kit, 100 reactions

• Ready to use format, including Master Mix for the target and PCR control to monitor for PCR inhibition and validate the quality
• Specific Primer and Probe mix for the pathogen/virus/viroid of interest
• Primer and Probe mix
• Positive and negative control to confirm the integrity of the kit reagents

HSV-1 TaqMan PCR Probe/Primer Set and Controls, 100 reactions

• Specific Primer/Probe mix and Positive Control for the pathogen/virus/viroid of interest
• Nuclease-free water
• Can be used together with Norgen’s PCR Master Mix (#28007) or customer supplied master mix

For research use only and NOT intended for in vitro diagnostics.

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. 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.

Component	Cat. TM32650 (100 preps)	Cat. TM32610 (100 preps)
MDx TaqMan 2X PCR Master Mix	2 x 700 μL	–
HSV-1 Primer & Probe Mix	280 μL	280 μL
HSV-1 Positive Control	150 μL	150 μL
Nuclease-Free Water (Negative Control)	1.25 mL	1.25 mL
Product Insert	1	1