12×20ml Angle Rotor 

Overview

• Rapid and simple procedure to generate digested peptides
• Simultaneous digestion, purification and concentration at once
• Peptide generation is complete, with no generation of additional artifacts being detected in mass spectrometry
• Peptides are ready for applications such as mass spectrometry and SDS-PAGE
• Purification is based on spin column chromatography that uses Norgen’s resin separation matrix

This kit is highly efficient in the enzymatic digestion of simple and complex protein samples using trypsin and the subsequent purification of the resulting peptides using a convenient spin column format.  Trypsin is added to the protein sample and bound to the column.  Salts are washed away and the trypsin is then activated to digest proteins.  Peptides are then eluted in a small volume and ready for downstream analysis. The peptides generated are complete, with no additional artifacts being detected in mass spectrometry.  Fifteen micrograms of protein can be processed, digested and purified with each spin column with about 20 minutes of hands-on time (plus trypsin incubation).   The simultaneous protein digestion and volumetric concentration of the purified peptides makes the kit a convenient method for preparing peptides to be analyzed by many downstream applications such as mass spectrometry and more.

Details

Supporting Data

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Kit Specifications
Maximum Protein Input	15 μg
Minimum Protein Input	2 μg
Minimum Elution Volume	30 μL
Time to Process 10 Purifications	20 minutes (Plus a 1-hour incubation)

Storage Conditions
All solutions should be kept tightly sealed and stored at room temperature. Once opened, the solution should be stored at 4°C. This kit is stable for 2 years after the date of shipment.

Component	Cat. 17500 (25 preps)
Wash Solution C	30 mL
Binding Buffer A	4 mL
Column Activation Buffer	3 mL
Micro Spin Columns	25
Collection Tubes	25
Elution tubes (1.7 mL)	25
Product Insert	1

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-V2 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V1-V2 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-V2 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-V2 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-V2 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. 70100 (24 preps)	Cat. 70110, 70120, 70130, 70140 (96 preps)
Amplicon PCR (PCR 1)	MGX Master Mix	330 µL	1,320 µL
	16S V1-V2 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

Propargyl-PEG3-Ms is a crosslinking reagent with a propargyl group and a mesyl group. The propargyl group reacts with azide-bearing compounds or biomolecules in copper catalyzed Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Propargyl-PEG3-Ms is a crosslinking reagent with a propargyl group and a mesyl group. The propargyl group reacts with azide-bearing compounds or biomolecules in copper catalyzed Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.