[DM3100] ExcelBand™ 1KB (0.25-10 kb) DNA Ladder, 500 μl

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 4 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 ITS2 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the fungal target ITS2 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 XPMagneticBeads(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 ITS2 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 ITS2 region of the fungal DNA. The post-PCR reaction is then cleaned up using AMPure XPbeads. 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 XPbeads. 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 ITS2 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 will remain stable for at least 1 year when stored at the specified storage conditions.

Introduction

Intended Use

For the selective isolation and culture of Listeria monocytogenes.

Principle and Interpretation

Peptone provides the carbon and nitrogen sources necessary for growth; yeast extract powder and starch provide carbon and nitrogen sources, vitamins and growth factors; sodium chloride can maintain a balanced osmotic pressure; glucose provides a carbon source; Listeria hydrolyzes esculin and reacts with iron ions to form black 6,7-dihydroxycoumarin; mannitol is a fermentable sugar, and phenol red is a pH indicator; lithium chloride and other antibiotics can inhibit Gram-negative bacteria and most Gram-positive bacteria; agar is the coagulant of the culture medium.

Formulation

Ingredients	/liter
Peptone	23 g
Starch	1 g
NaCl	5 g
Columbia agar	13 g
Mannitol	10 g
Ferric ammonium citrate	0.5 g
Esculin (aesculin)	0.8 g
Dextrose (glucose)	0.5 g
Lithium chloride	15.0 g
Phenol red	0.08 g
pH7.2±0.2 at 25°C

Preparation

Weigh 68.9g of dry powder of this product, add 1L of distilled water or deionized water, stir, heat and boil until completely dissolved, divide into Erlenmeyer bottles, sterilize at 121℃ for 15min, cool to room temperature and set aside. Heat to dissolve agar before use, cool to 50℃, add 1 tube of supporting reagent (SR0140) per 100mL of basal culture medium, shake well and pour into sterilized culture dish.

Quality Control

The following quality control strains were inoculated and cultured at 35-37℃ for 24h. The results are as follows:

Quality control strains	Growth
Listeria monocytogenes ATCC19115	Gray-green colonies with a black depression in the center and black surrounding
Enterococcus faecalis ATCC29212	–
Escherichia coli ATCC25922	–

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.

Intended Use For the selective isolation and culture of Listeria monocytogenes. Principle and Interpretation Peptone provides the carbon and nitrogen sources necessary for growth; yeast ex……