Propargyl-PEG9-bromide

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

This product is suitable for extracting total RNA from 50-150mg conventional plantor fungal samples as well as samples rich in polyphenolic and polysaccharides. This kit is based on silica gel column purification technology, and the extraction process does not require the use of toxic phenol chloroform extraction. The entire extraction process only takes 20-30 minutes. This kit adopts DNA filtration technology, which can efficiently filter and remove DNA. The obtained RNA can be directly used for experiments such as RT-PCR, Northern Blot, poly A+ purification, nucleic acid protection, and in vitro translation.

Details

Kit Contents

Contents	R415002	D415003
Purification Times	50 Preps	250 Preps
HiPure RNA Mini Columns	50	250
gDNA Filter Columns	50	250
2ml Collection Tubes	100	500
TCEP (1M)	0.29 g	5 x 0.29 g
Buffer EP	1.0 ml	5.0 ml
Buffer PSL	50 ml	250 ml
Buffer RW1	50 ml	250 ml
Buffer RW2*	20 ml	2 x 50 ml
RNase Free Water	10 ml	30 ml

Storage and Stability

Except TCEP, the kit components can be stored dry at room temperature (15–25°C) and are stable for at least 18 months under these conditions. After receiving the product, it is recommended to store TCEP (dry powder) at -20-8°C. At low temperatures, Buffer PSL may form precipitates, dissolve it completely by 55°C water bath.

Purchase Guide

1. When dealing with woody or uncommon samples, R4150 is recommended first. R4150 contains two polysaccharide/polyphenol lysis buffer, which is the most universal product.

2. R4151 is recommended for handling common economic crop samples for the first time. Strong lysis solution can be used to process easy-extraction samples. The amount of corn or rice leaves samples can reach up to 300mg.

3. R4165 adopts CTAB/chloroform method, which can also handle a large number of difficult-to-extraction plants, but requires contact with chloroform substitutes, which is less safe than other kits. This kit uses DNase Ⅰ to remove DNA, which is also a good choice for extracting polysaccharide/polyphenol-rich plant samples.

4. R4014 is recommended for fruit/starch plant samples, which uses improved trizol pre-treatment, single column operation and is more economical.

This product is suitable for extracting total RNA from 50-150mg conventional plantor fungal samples as well as samples rich in polyphenolic and polysaccharides. This kit is based on silica gel column purification technology, and the extraction process does not require the use of toxic phenol chloroform extraction. The entire extraction process only takes 20-30 minutes. This kit adopts DNA filtration technology, which can efficiently filter and remove DNA. The obtained RNA can be directly used for experiments such as RT-PCR, Northern Blot, poly A+ purification, nucleic acid protection, and in vitro translation.

Introduction

Usages:
For cultivating, enumerating and preserving nonfastidious bacteria.

Principle:
Peptone and beef extract powder to provide nitrogen, vitamins, amino acids and carbon; agar as medium coagulant.

Formulation(per liter):
Peptone: 5g
Beef extract :3g
Agar :15g
Final pH 6.8 ± 0.2

How to use:
1. Suspend 23g of product, adding 1L of distilled or deionized water, heated to boiling stirring until completely dissolved, dispensing into flask, autoclave at 121 ℃ for 15min, set aside.
2.Diluted and treated samples.

Quality control：
          　　

Item	The name and number of strain	Growth	Colony Color
1	Escherichia coli ATCC25922	Good	Dark red
2	Aerogenes CMCC (B) 45103	Good	red
3	Staphylococcus aureus ATCC6538	Suppressed	—

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

Specifications: 250g/bottle

250g