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ITS2 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 4 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 ITS2 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the fungal target ITS2 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 XPMagneticBeads(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 ITS2 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 ITS2 region of the fungal DNA. The post-PCR reaction is then cleaned up using AMPure XPbeads. 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 XPbeads. The libraries are then ready for quantification, pooling and sequencing.16S Library Prep

Details

Supporting Data

Figure 1 / 3

Previous

Figure 1. Amplicons generated by ITS2 amplicon primers, cleaned and run on a 2% agarose gel.
Figure 1. Amplicons generated by ITS2 amplicon primers, cleaned and run on a 2% agarose gel.
Figure 2. Libraries generated by indexing PCR, cleaned and run on a 2% agarose gel.
Figure 3. Bioanalyzer traces of ITS2 libraries generated by indexing PCR, cleaned and run on a high sensitivity DNA chip.

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 ITS2 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 will remain stable for at least 1 year when stored at the specified storage conditions.

Other Products

2400ml Bottle

X96 Dual Ring Magnetic Plate 96-well

Permagen’s most universal ring magnet plate. From Low elution PCR applications up to 2 mL deep well, the X96 has you covered. No need to purchase two separate plates anymore. Smaller inner ring magnet allows for volumes as low as 5 µL, larger outer magnet handles up to 2 mL deep well 

ANSI/ SBS Footprint (127.75mm x 85.50mm) to fit into any automated liquid handling robot on bottom, SBS/ SLAS fit on top to accept any microplate

Integrated Cushion base for maximum recovery. Helps aid in set-up, robot positioning inconsistencies, and labware consumable differences 

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

SKU

A011623

A011623KT (Kit for 0.2 ml tubes)

Features

  • ANSI/ SBS footprint
  • Dual ring magnet technology 
  • Cushion base for added benefits
  • Solid Aluminum alloy design
  • Precision manufactured for more consistent results
  • Fast magnetic bead separations
  • Made in USA 
  • For Research use only

Compatibility

  • Round bottom plates 
  • Pyramid bottom plates 
  • Flat Bottom plates
  • Deep well plates
  • Full-, Semi-, and un-Skirted PCR Plates
  • Any magnetic beads
  • Any common liquid handlers i.e. Tecan®, Opentrons®, Hamilton®, Agilent®, Beckman®, etc.

Volumes

Maximum – 2 mL (Deep Well)

Minimum – 5 µL (PCR Plate)

MZ45 Single Cylinder Series Zoom Lens

Specification

Optical Microscope0.7-4.5X zoom objective
10X Eyepiece,view by eyepiece together
Working Distance100mm
Display8 inch color monitor,resolution 640*480,OSD menu,with cross-line function
Digital Camera1/3 Sensor chip,80OTVL(PAL)OSD menuO
Output InterfaceHDMI,VGA Interface
Magnification12.6-81X(customized magnification is available according to customers’need)O
AccessoriesDC 12V 2A power supply,USB cable,CD-ROM
ModelMZ45D
Opticalmagnification0.7X—5X
Zoom Ratio1:7
EyepieceStandard 0.4X
Option1X
Objective0.3X,0.5X,0.75X,1X,1.5X,2X
Video SystemOptional 1/2,1/3 CCD digital camera
WorkingDistance100mm
Accessorieslllumination
Various Stands
ModelMZ45
Opticalmagnification0.7X—5X
Zoom Ratio1:7
EyepieceStandard 0.4X
Option1X
Objective0.3X,0.5X,0.75X,1X,1.5X,2X
Video SystemOptional 1/2,1/3 CCD digital camera
WorkingDistance00mm
Accessoriesumination
Various Stands