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16S V1-V3 Library Preparation Kit for Illumina

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Detail

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.16S Library Prep

Details

Supporting Data

Figure 1 / 3

Previous

Figure 1: 16S V1-V3 PCR1 Amplification
Figure 1: 16S V1-V3 PCR1 Amplification
Figure 2: 16S V1-V3 Indexed Libraries
Figure 3: BioAnalyzer Trace of the 16S V1-V3 Indexed Library

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 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.

StepComponentCat. 70200 (24 preps)Cat. 70210, 70220, 70230, 70240 (96 preps)
Amplicon PCR (PCR 1)MGX Master Mix330 µL1,320 µL
16S V1-V3 Primer Mix70 µL280 µL
Index PCR (PCR 2)Indexing Master Mix660 µL2 x 1,320 µL
N7xx Index Primer50 µL50 µL
S5xx Index Primer70 µL70 µL
PCR Clean-UpResuspension Buffer2 x 1,250 µL2 x 5,000 µL
Nuclease-free water1,250 µL1 x 6,000 µL

Other Products

Propargyl-PEG4-thiol

Propargyl-PEG4-thiol is a crosslinker containing a propargyl group and thiol group. The propargyl group can form triazole linkage with azide-bearing compounds or biomolecules via copper catalyzed Click Chemistry reactions. The thiol group reacts with maleimide, OPSS, vinylsulfone and transition metal surfaces including gold, silver, etc. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

96-Well Low Elution Ring Magnetic Plate

Designed for those labs wishing to separate magnetic beads into rings.

The Permagen ring magnet low elution plate was designed for use in automation applications where volumes as low as 5 µL are required. Most PCR plates are bent, leading to inconsistent lab results. Unlike most products, we have added an angled frame around the top of our plate, this helps with two things, straightening out the PCR plate, and leading it to the proper location on the magnets during automation

SBS SLAS Footprint (127.75mm x 85.50mm) to fit into any automated liquid handling robot 

Features include solid aluminum alloy construction and hard coat anodized finish for years of trouble-free use, and compatible with any magnetic beads

SKU #

RLE500

Features

  • SBS/ SLAS footprint
  • Solid Aluminum alloy design
  • Precision manufactured for more consistent results
  • Fast magnetic bead separations
  • Integrated bracket to help keep microplate in place and flat
  • Made in USA 
  • For Research use only

Compatibility

  • Full- skirt PCR plates
  • 1/2- skirt PCR plates 
  • Un-skirted PCR plates
  • Any magnetic beads
  • Any common liquid handlers i.e. SPTLabtech Firefly®, Tecan®, Opentrons®, Hamilton®, Agilent®, Beckman®, etc.

Volumes

Maximum  – 150 µL 

Minimum  – 5 µL 

t-Boc-aminooxy-PEG4-propargyl

t-Boc-aminooxy-PEG4-propargyl is a click chemistry tool containing a propargyl group and t-Boc-aminooxy group. Propargyl group is reactive with azide-bearing compounds or biomolecules via copper catalyzed Click Chemistry to yield a stable triazole linkage. T-Boc-aminooxy can be deprotected under mild acidic conditions and then can react with an aldehyde or ketone group to form a linkage. The hydrophilic PEG spacer increases solubility in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.