Methyltetrazine-PEG3-DBCO

Introduction

Usages:
For cultivation of bifidobacteria and lactic acid metabolites test.

Principle:
Peptone, yeast extract provide nitrogen, vitamins, and growth factors; glucose as
fermentable sugars; cysteine – HCl, vitamin K1, hemin, magnesium sulfate, calcium
chloride, sodium chloride, sodium bicarbonate training offers various bifidobacteria growth factor; dipotassium hydrogenphosphate, potassium dihydrogenphosphate buffer.

Formulation ( per liter):
Peptone 20g
Glucose 5g
Yeast powder 10g
Cysteine – hydrochloride 0.5g
Calcium chloride 0.008g
Magnesium sulfate 0.008g
Dipotassium hydrogen phosphate 0.04g
Potassium dihydrogen phosphate 0.04g
Sodium bicarbonate 0.4g
Sodium chloride 0.08g
Final pH 6.0 ± 0.2

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

250g

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

Description 

The ExcelDye™ 6× DNA Loading Dye (Green) is pre-mixed buffer for tracking the DNA sample during the electrophoresis on agarose or polyacrylamide gels. It contains two dyes (Xylene cyanol FF and Orange G) for tracking the DNA migration. The Xylene cyanol FF and Orange G migrate at approximately 800 bp and 30 bp on a standard 2% TAE agarose gel respectively (4,000 bp and 50 bp on 1% TAE agarose gel respectively). The included glycerol keeps the DNA at the bottom of the well and the presence of EDTA chelates divalent metal ions to prevent the process of metal-dependent nuclease.

Composition 

• 0.03% Xylene cyanol FF 
• 0.15% Orange G 
• 10 mM Tris-HCl (pH 8.0) 
• 60% glycerol 
• 60 mM EDTA

Storage 

4°C for 12 months
-20°C for 36 months

The ExcelDye™ 6× DNA Loading Dye (Green) is pre-mixed buffer for tracking the DNA sample during the electrophoresis on agarose or polyacrylamide gels. It contains two dyes (Xylene cyanol FF and Orange G) for tracking the DNA migration. The Xylene cyanol FF and Orange G migrate at approximately 800 bp and 30 bp on a standard 2% TAE agarose gel respectively (4,000 bp and 50 bp on 1% TAE agarose gel respectively). The included glycerol keeps the DNA at the bottom of the well and the presence of EDTA chelates divalent metal ions to prevent the process of metal-dependent nuclease.