Bis-PEG11-DBCO

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

Figure 1 / 3

Previous

Next

Click for expanded view

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 ExcelRT™ One-Step RT-PCR Kit is designed for the reverse transcription and PCR amplification of a specific target RNA from either total RNA or mRNA. The ExcelRT™ One-Step RT-PCR Kit provides the user an alternative to the lengthy two step processes (first strand generation and amplification) by using a single mixture, single tube, one step reaction. The ExcelRT™ One-Step RT-PCR Kit contains a 2X reaction premix consisting of an optimized buffer, dNTPs, Mg²⁺ and enzyme stabilizer, and a blend of recombinant reverse transcriptase and Taq DNA polymerase. The ExcelRT™ One-Step RT-PCR Kit allows the user to complete the RT-PCR process using a thermocycler in a single reaction setting. The ExcelRT™ One-Step RT-PCR Kit is capable of detecting even trace amounts of target RNA and ideal for target RNA amplification and analysis. 

Features

• Generates dsDNA directly from RNA samples 
• High sensitivity and yields 
• High reproducibility, reduced pipetting errors
• High throughput 
• Reduced RNase H ribonuclease activity

Storage

-20°C for 24 months 

Contents

High sensitivity and yields 

The ExcelRT™ One-Step RT-PCR Kit is designed for the reverse transcription and PCR amplification of a specific target RNA from either total RNA or mRNA. The ExcelRT™ One-Step RT-PCR Kit provides the user an alternative to the lengthy two step processes (first strand generation and amplification) by using a single mixture, single tube, one step reaction. The ExcelRT™ One-Step RT-PCR Kit contains a 2X reaction premix consisting of an optimized buffer, dNTPs, Mg2+ and enzyme stabilizer, and a blend of recombinant reverse transcriptase and Taq DNA polymerase. The ExcelRT™ One-Step RT-PCR Kit allows the user to complete the RT-PCR process using a thermocycler in a single reaction setting. The ExcelRT™ One-Step RT-PCR Kit is capable of detecting even trace amounts of target RNA and ideal for target RNA amplification and analysis.

Overview

• Ready-to-use
• Quantitative
• Highly Stable
• Precise
• Ten discrete fragments ranging from 300 bp to 5000 bp

The Norgen MidRanger 1 kb DNA Ladder is prepared to ensure quality and batch-to-batch consistency. Our MidRanger contains ten discrete fragments ranging from 300 bp to 5000 bp. This Ladder is ideal for sizing larger PCR products (>1kb) and most cloning applications.

Contents:
1mL of premixed DNA ladder (0.5µg/10µL) in loading buffer (10mM EDTA, 10% glycerol, 0.015% bromophenol blue, and 0.17% SDS).

Details

Supporting Data

Figure 1 / 1

Previous

Next

Click for expanded view

MidRanger 1kb DNA Ladder (Cat# 11700) – 100 loads

Ladder Properties:
• Ten discrete fragments ranging from 300 bp to 5000 bp

Fragment	Size (bp)	Mass (ng)
1	5000	88
2	4000	78
3	3000	67
4	2500	58
5	2000	50
6	1500	43
7	1000	25
8	700	34
9	500	33
10	300	24

Recommended Use:

Mix thoroughly. For best results, load 10µL of DNA ladder per well. For precise mass determination with a densitometer, stain gel after electrophoresis using 0.5µg/mL ethidium bromide for 30-40 minutes. The table above shows the size and mass for each band based on 10µL ladder per well.

Storage: 

Stable at room temperature. For longer term storage, -20°C is recommended.

This ladder was standardized using 10µL of DNA per lane on a 0.8 cm thick, 13 x 15 cm, 1.0% agarose gel run in TAE buffer.