GeneAb™ PD-L1

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 V1-V3 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V1-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 V1-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 V1-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 V1-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. 70200 (24 preps)	Cat. 70210, 70220, 70230, 70240 (96 preps)
Amplicon PCR (PCR 1)	MGX Master Mix	330 µL	1,320 µL
	16S V1-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

Introduction

Usages:
For selective cultivation of bacteria especially Vibrio.

Principle:
Peptone provide carbon and nitrogen sources; sodium chloride to maintain osmotic equilibrium; higher pH can suppress the growth of coliform and other bacteria, it is conducive to the growth of Vibrio cholerae.

Formulation（per liter）:
Peptone 10g
Sodium chloride 10g
Final pH 8.5 ± 0.2

How to use：
1.Suspend 18g in 1L of distilled water, stirring heated to boiling to completely dissolve, autoclave at 121℃ for 15 minutes.
2.Diluted and treated samples.

Quality control：
          　　

Item	The name and number of strain	Growth	Colony Color
1	Vibrio cholerae VbO	Good	Turbid broth
2	Escherichia coli ATCC25922	Poor	—

Storage: Keep container tightly closed, store in a cool, dry place, away from bright light. Storage period of 3 years.

Specifications: 250g/bottle 

250g

DBCO-NHCO-PEG5-NHS ester is a PEG linker containing NHS ester that is able to react specifically and efficiently with primary amines such as the side chain of lysine residues or aminosilane-coated surfaces at neutral or slightly basic condition to form a covalent bond. The hydrophilic PEG spacer arm improves water solubility and can also provide a long and flexible connection that minimizes steric hindrance during ligation. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

DBCO-NHCO-PEG5-NHS ester is a PEG linker containing NHS ester that is able to react specifically and efficiently with primary amines such as the side chain of lysine residues or aminosilane-coated surfaces at neutral or slightly basic condition to form a covalent bond. The hydrophilic PEG spacer arm improves water solubility and can also provide a long and flexible connection that minimizes steric hindrance during ligation. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.