m-PEG2-DBCO

Introduction

Intended Use

For the cultivation of E. coli in fermentation and molecular genetic studies.

Principle and Interpretation

Peptone provides nitrogen . Vitamins (including B vitamins) and certain trace elements are provided by yeast extract. Glucose provides carbon.Sodium ions for transport and osmotic balance are provided by sodium chloride. Agar is the solidifying agent.

Formulation

Ingredients	/liter
Peptone	10.0g
Yeast extract	5.0g
Sodium chloride	5.0g
Glucose	1.0g
Agar	15.0g
pH7.0±0.2 at 25°C

Preparation

Suspend 36g in 1 litre of distilled water. Bring to a boil to dissolve completely. Sterilize by autoclaving at 121°C for 15minutes.

Quality Control

Cultural characteristics observed after incubation at 35-37°C for 24 hours

Quality control strains	Growth
Escherichia coli  ATCC25922	good

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.

Propargyl-PEG6-amine is a crosslinker with a hydrophilic PEG spacer which can increase the hydrophilicity of the molecule in aqueous environment. The amine group is reactive with carboxylic acids, activated NHS esters, carbonyls (ketone, aldehyde) etc. The propargyl group can react with azide-bearing compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Propargyl-PEG6-amine is a crosslinker with a hydrophilic PEG spacer which can increase the hydrophilicity of the molecule in aqueous environment. The amine group is reactive with carboxylic acids, activated NHS esters, carbonyls (ketone, aldehyde) etc. The propargyl group can react with azide-bearing compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

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