022010P1 Nutrient Broth

Description 

The DM2360 FluoroBand™ 100 bp+3K Fluorescent DNA Ladder is a ready-to-use DNA ladder, which is pre-mixed with high sensitivity DNA binding fluorescent dye and loading dye for direct gel loading. The DNA Ladder DM2360 is composed of 12 individual DNA fragments: 3k, 1.5k, 1k, 900, 800, 700, 600, 500, 400, 300, 200 and 100 bp derived from a mixture of PCR products and specifically digested plasmid DNA; these bands can be visualized when illuminated with 470 nm blue light or UV light. This product contains two enhanced bands (1.5 kb and 500 bp) for easier reference. In addition, two tracking dyes, Xylene cyanol FF and Orange G which mimic the migration of 4,000 bp and 50 bp dsDNA during electrophoresis are also added for real time monitoring. Real time observation of the electrophoresis is also possible if compatible light source is fitted to the electrophoresis tank. 

Features

• Sharp bands
• Quick reference— enhanced bands 
• Ready-to-use— premixed with loading dye for direct loading
• Stable— room temperature storage over 6 months
• Directly observed by UV or blue light— premixed with high sensitive DNA fluorescent dye

Source 

Phenol extracted PCR products and dsDNA digested with specific restriction enzymes, equilibrated in 10 mM Tris-HCl (pH 8.0) and 10 mM EDTA.

Range 

100 ~ 3,000 bp 

Concentration 

56 µg/ 500 µl

Recommended loading volume 

5 µl/ well

Storage

Protected from light 
Room temperature for 6 months
4°C for 12 months 
-20°C for 24 months 

Description

Salmonella spp. are members of the family Enterobacteriaceae. They are Gram-negative, facultatively anaerobic, flagellated, rod-shaped organisms. They are approximately 0.7 to 1.5 µm in diameter and 2 to 5 µm in length and responsible for a large number of cases of foodborne illness throughout the world. Salmonella have circular DNA genomes with a mean length of approximately 4530 kb, although this can vary by up 1000 kb. Salmonella classification is extremely complex, however, the genus is divided into two species: S. enterica and S.bongori. S. enterica is then itself divided into 6 biochemically distinct subspecies and the Salmonella genus is further classified into serovars (serotypes) based on the lipopolysaccharide (O), flagella protein (H), and sometimes the capsular (VI) antigens. There are more than 2500 known serovars and within a serovar there may be strains that differ in virulence.

Salmonella are mainly transmitted by the faecal-oral route. They are carried asymptomatically in the intestines or gall bladder of many animals, being continuously or intermittently shed in the faeces. Humans can become infected if they do not wash their hands after contact with infected animals or animal faeces. In such instances the bacteria adhere to and enter the cells of the intestinal epithelium. The toxins produced by the bacteria can damage and kill the cells that line the intestines, which results in intestinal fluid loss. The bacteria can survive for weeks in a dry environment and far longer in water thus they are frequently present in polluted waters. Salmonella can also be carried latently in the mesenteric lymph nodes or tonsils; these bacteria are not shed, but can become reactivated after stress or immunosuppression. In addition, fomites and vectors can spread Salmonella and vertical transmission occurs in birds, with contamination of the vitalize membrane, albumen and possibly the yolk of eggs. Salmonella spp. can also be transmitted in utero in mammals.

There are two different disease conditions that are distinct to salmonellosis; gastroenteritis and enteric typhoid fever. The gastroenteritis is a nonsystemic infection of the intestinal tract and regional lymph nodes that gives rise to headache, muscle aches, diarrhoea, vomiting, abdominal cramping, chills, fever, nausea and dehydration. In contrast, the enteric typhoid fever is a systemic disease in which the microorganism replicates within the cells of the reticuloendothelial system. The symptoms usually appear 6 to 72 hours after ingesting contaminated food although individuals can be infected with the bacteria without having symptoms. Those with and without symptoms shed the bacteria in their stool and it is important that personal hygiene be maintained at all times.

Overview

• Protocol optimized for DNA isolated from a diversity of samples including stool, soil, water, saliva, plant, urine, skin, and more
• Simple and quick workflow: library could be prepared in less than 5 hours
• Component of Norgen’s metagenomics workflow
• A single NGS run can be prepared with up to 384 unique dual-index libraries

Sample type purification kit guide 

The 16S V2-V3 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V2-V3 amplicon libraries to be used for next-generation sequencing on Illumina platforms. All molecular reagents including primers, enzyme mixes, indexes, and buffers are provided. Instructions for PCR clean up with the AMPure XP Magnetic Beads (supplied by customer) are also included for rapid purification of nucleic acid products generated at two steps of the workflow. The library prep workflow could be used for purified DNA inputs from different sources including stool, soil, water, saliva, plant, urine, skin swab, vaginal swab, cheek swab, nasal swab, plasma/serum, tongue swab, gum swab, and others.

The 16S V2-V3 Library Preparation Kit for Illumina has a streamlined procedure that reduces the handling time such that the library prep procedure can be completed in approximately 4 hours (see diagram below). Input DNA is first subjected to targeted PCR to amplify the V2-V3 region of the DNA encoding 16S rRNA. The post-PCR reaction is then cleaned up using AMPure XP beads. Dual index primers are then added using a limited-cycle PCR. The indexed amplicons flanked by 5′ and 3′ barcoded adaptors are then cleaned using AMPure XP beads. The libraries are then ready for quantification, pooling and sequencing.

Details

Supporting Data

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Minimum amount of starting material:	2.5 µL of DNA (5 ng/µL)
Time to complete library preparation:	4 hours

Storage Conditions and Product Stability
Norgen’s 16S V2-V3 Library Prep Kit for Illumina is shipped as one kit box (for the 24 prep kit) or two sub-component kits (for the 96 prep kit). All kits should be stored at -20°C upon arrival.

All kit components should remain stable for at least 1 year when stored at the specified storage conditions.

Step	Component	Cat. 70300 (24 preps)	Cat. 70310, 70320, 70330, 70340 (96 preps)
Amplicon PCR (PCR 1)	MGX Master Mix	330 µL	1,320 µL
	16S V2-V3 Primer Mix	70 µL	280 µL
Index PCR (PCR 2)	Indexing Master Mix	660 µL	2 x 1,320 µL
	N7xx Index Primer	50 µL	50 µL
	S5xx Index Primer	70 µL	70 µL
PCR Clean-Up	Resuspension Buffer	2 x 1,250 µL	2 x 5,000 µL
	Nuclease-free water	1,250 µL	1 x 6,000 µL