[QP4510] Q-PAGE™ TGN Precast Gel (Mini, 12 wells, 4-15%), 10 gels

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

Description 

The DM3200 ExcelBand™ 1 KB Plus (0.1-10 kb) DNA Ladder is a ready-to-use DNA ladder, which is pre-mixed with loading dye for direct gel loading. The DNA Ladder DM3200 is composed of 19 individual DNA fragments: 10k, 8k, 6k, 5k, 4k, 3k, 2.5k, 2k, 1.5k, 1k, 900, 800, 700, 600, 500, 400, 300, 200, and 100 bp derived from a mixture of PCR products and specifically digested plasmid DNA. This product contains three enhanced bands (3 kb, 1.5 kb and 500 bp) for easier reference. In addition, three tracking dyes, Xylene cyanol FF, Bromophenol blue and Orange G which mimic the migration of 4,000 bp, 500 bp and 50 bp dsDNA during electrophoresis are also added for real time monitoring. 

Features

• Sharp bands
• Quick reference— enhanced bands 
• Ready-to-use— premixed with loading dye for direct loading
• Stable— room temperature storage over 6 months

Source 

Phenol extracted PCR products and dsDNA digested with specific restriction enzymes, equilibrated in 10 mM Tris-HCl (pH 8.0) and 10 mM EDTA. 

Range 

100 ~ 10,000 bp 

Concentration 

87 µg/ 500 µl 

Recommended loading volume 

5 µl/ well 

Storage

Room temperature for 6 months
4°C for 12 months 
-20°C for 36 months

The DM3200 ExcelBand™ 1 KB Plus (0.1-10 kb) DNA Ladder is a ready-to-use DNA ladder, which is pre-mixed with loading dye for direct gel loading. The DNA Ladder DM3200 is composed of 19 individual DNA fragments: 10k, 8k, 6k, 5k, 4k, 3k, 2.5k, 2k, 1.5k, 1k, 900, 800, 700, 600, 500, 400, 300, 200, and 100 bp derived from a mixture of PCR products and specifically digested plasmid DNA. This product contains three enhanced bands (3 kb, 1.5 kb and 500 bp) for easier reference. In addition, three tracking dyes, Xylene cyanol FF, Bromophenol blue and Orange G which mimic the migration of 4,000 bp, 500 bp and 50 bp dsDNA during electrophoresis are also added for real time monitoring.

Propargyl-PEG9-alcohol is a crosslinker that can participate in copper catalyzed azide-alkyne Click Chemistry reactions to form stable triazole linkage. The PEG spacer increases the hydrophilicity of the molecule in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Propargyl-PEG9-alcohol is a crosslinker that can participate in copper catalyzed azide-alkyne Click Chemistry reactions to form stable triazole linkage. The PEG spacer increases the hydrophilicity of the molecule in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.