Overview

• Detection kits for Campylobacter jejuni
• Available in TaqMan format for analysis.

Campylobacter jejuni is a curved, rod-shaped and microaerophillic gram negative bacterium. It is one of the most common causal agents of gastroenteritis with diarrhea as the main symptom. While infection of C. jejuni is seldom life-threatening, it is considered one of the most common food-borne bacteria with over 2 million people infected per year in US alone. Infection of C. jejuni usually results from consumption of poorly prepared food including undercooked meat (particularly poultry), untreated water or raw unpasteurized milk. Traditional identification of C. jejuni involves culturing, however the microaerophilic characteristic of this bacterium make the enrichment process laborious and costly.

Campylobacter jejuni 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

Campylobacter jejuni 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. TM36150 (100 preps)	Cat. TM36110 (100 preps)
MDx TaqMan 2X PCR Master Mix	2 x 700 μL	–
Campylobacter jejuni Primer & Probe Mix	280 μL	280 μL
Campylobacter jejuni Positive Control	150 μL	150 μL
Nuclease-Free Water (Negative Control)	1.25 mL	1.25 mL
Product Insert	1	1

DBCO-PEG4-Desthiobiotin is a PEG linker containing a desthiobiotin group and a DBCO functional group. Desthiobiotin is used for affinity-based applications such as pull-down assays or for ligating with streptavidin proteins while DBCO is a click chemistry handle that quickly reacts with azide groups on target molecules. Desthiobiotin is a sulfur-free analogue of biotin which binds streptavidin with slightly less strength than biotin, which provides it with a soft-release characteristic that is useful for in pull-down assays by minimizing co-elution with endogenous biotinylated molecules. The inclusion of a PEG linker in this molecule improves its aqueous solubility.

DBCO-PEG4-Desthiobiotin is a PEG linker containing a desthiobiotin group and a DBCO functional group. Desthiobiotin is used for affinity-based applications such as pull-down assays or for ligating with streptavidin proteins while DBCO is a click chemistry handle that quickly reacts with azide groups on target molecules. Desthiobiotin is a sulfur-free analogue of biotin which binds streptavidin with slightly less strength than biotin, which provides it with a soft-release characteristic that is useful for in pull-down assays by minimizing co-elution with endogenous biotinylated molecules. The inclusion of a PEG linker in this molecule improves its aqueous solubility.

Introduction

Free-circulating nucleic acids, such as tumor-specific extracellular DNA fragments and mRNAs in the blood or fetal nucleic acids in maternal blood, are present in serum or plasma usually as short fragments, <1000bp (DNA). HiPure Circulating DNA Midi Kit enables efficient purification of these circulating nucleic acids from human plasma, serum, or urine. The extracted products can be used for clinical in vitro detection.

Details

Specifications

Features	Specifications
Main Functions	Isolation circulating DNA from 1-5ml plasma, serum, body fluids using vacuum protocol
Applications	qPCR, liquid or solid chip analysis, hybridization and SNP detection, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (vacuum)
Sample type	Serum, plasma and other cell-free fluid samples
Sample amount	1-5ml
Elution volume	≥50μl
Time per run	≤60 minutes
Liquid carrying volume per column	4ml
Binding yield of column	1mg

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.

Advantages

• High yield – most optimal process, free DNA (>50bp) can be obtained to the maximum extent
• High concentration – low elution volume, ensuring high nucleic acid concentration
• High purity – low alcohol binding method, completely removing inhibitor and protein pollution
• High recovery – DNA can be recovered at thelevel of PG by silica gel column purification

Kit Contents

Contents	IVD3182
Purification Times	50
Buffer ACL	250 ml
Buffer ACB*	300 ml
Buffer DCW1*	22 ml
Buffer DCW2*	10 ml
Proteinase K	540 mg
Protease Dissolve Buffer	30 ml
Carrier RNA	110 μg
Nuclease Free Water	20 ml
HiPure CFDNA Mini Columns	50
2 ml Collection Tubes	100
Extender Tube	50
Vac-Connector	50

Storage and Stability

Proteinase K, Carrier RNA 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 dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. The entire kit can be stored at 2-8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.

Experiment Data

Free-circulating nucleic acids, such as tumor-specific extracellular DNA fragments and mRNAs in the blood or fetal nucleic acids in maternal blood, are present in serum or plasma usually as short fragments,