AgriSeq™ HTS Library Kit
AgriSeq™ HTS Library Kit
Applied Biosystems™

AgriSeq™ HTS Library Kit

The Applied Biosystems AgriSeq HTS Library Kit is specifically designed for high-throughput preparation of amplicon libraries for targeted genotyping-by-sequencing (GBS)Read more
Have Questions?
Change viewbuttonViewtableView
Catalog NumberNo. of Reactions
A341439600 Reactions
A34144960 Reactions
Catalog number A34143
Price (CNY)
-
No. of Reactions:
9600 Reactions
The Applied Biosystems AgriSeq HTS Library Kit is specifically designed for high-throughput preparation of amplicon libraries for targeted genotyping-by-sequencing (GBS) applications in agrigenomics.

Key product features include:
• Exceptional uniformity of multiplexed target amplification generates robust, reproducible results
• Simplified workflow enables high-throughput sample processing
• Convenient configuration contains library normalization reagents and volumes sufficient for automated platforms

Excellent data quality
AgriSeq library preparation has been optimized for targeted GBS of panels ranging from 100–5000 amplicons. With a highly efficient and specific multiplex technology, AgriSeq amplification can generate libraries with high uniformity (87.2–99.3%), percent on-target reads (85.7–99.7%), and overall genotype call rates (87.5–99.7%) based on a set of nine recently designed panels ranging in size from 62–3000 amplicons across seven different species. The figure below shows an average of 98.2% uniformity for a 3000-amplicon panel across 192 samples (i.e., 98% of bases are covered by at least 0.2X of the average read depth). Even coverage is significant in that it reduces the amount of sequencing required to achieve consistent genotype calls, thus reducing the sequencing cost per sample. In comparison, weaker multiplexing performance is reported with the competitor technology TruSeq, which provides only 88% average uniformity and a reported 1536 maximum multiplexing capability (see reference).

High throughput sample processing
The AgriSeq Library workflow (see figure below) can be performed in either 96- or 384-well plates for high-throughput sample processing using as little as 10 ng of DNA input per reaction. The workflow has been optimized to minimize sample handling and ancillary reagent usage through the processing of pooled samples. This allows the workflow to be completed with minimal pipetting steps, helping to reduce the potential for error, as well as simplifying automation setup on common third party platforms such as Tecan or Hamilton systems. Standard data analysis processing includes automated quality assessment, alignment, and genotype calling using the included Torrent Suite software and Variant Caller plug-in without the need for a bioinformatician, expensive data storage, or third party programs.

Conveniently packaged containing library normalization reagents & volumes for automation
The AgriSeq HTS Kit is offered in large pack sizes (960 or 9600 reactions) to accommodate high throughput agrigenomics projects while minimizing the cost and waste of unnecessary packaging. The AgriSeq HTS Kit includes not only the reagents for library preparation, but all of the components required for library normalization to help achieve consistent results between samples, and sufficient overage has been provided to accommodate automated workflows. With a low price per sample exclusively available for agricultural customers, AgriSeq targeted GBS is an economical way to generate high-throughput genotyping data to advance your breeding programs.

For Research Use Only. Not for use in diagnostic procedures.
Specifications
For Use With (Application)Sequencing
For Use With (Equipment)Ion GeneStudio S5 System
LibrariesTargeted Sequencing Library
No. of Reactions9600 Reactions
Product LineAgriSeq
Product TypeHTS Library Kit
Quantity9600 reactions
Sample TypeDNA
Shipping ConditionApproved for shipment at Room Temperature or on Wet or Dry Ice
Workflow StepLibrary Generation
FormatKit
Sequencing TypeGenome and DNA Sequencing
Unit SizeEach
Contents & Storage
Store in freezer (-5 to -30°C).

Frequently asked questions (FAQs)

With the AgriSeq HTS Library Kit, I see high molecular weight material present during sequencing. Why is this?

Here are possible causes and recommended solutions:
-High molecular weight DNA may not have been removed during purification of the amplified library. No action is required.
-Inserts may have formed concatemers during the ligation step. We recommend reducing the nucleic acid input, requantifying using the Qubit 1X dsDNA HS Assay Kit, Qubit dsDNA HS Assay Kit, or the Quant-iT DNA Assay Kit, High Sensitivity and Qubit Fluorometer, and reducing the target amplification cycle number.
Note: High molecular weight DNA does not interfere with sequencing.

Find additional tips, troubleshooting help, and resources within our Capillary Electrophoresis Applications Support Center.

Why is my library concentration high with the AgriSeq HTS Library Kit?

Here are possible causes and recommended solutions:
-The sample DNA may have been misquantified. We recommend requantifying using the Qubit 1X dsDNA HS Assay Kit, Qubit dsDNA HS Assay Kit, or the Quant-iT DNA Assay Kit, High Sensitivity and Qubit Fluorometer.
-More than 10 ng of sample DNA may have been used. We recommend adding the recommended amount of DNA (10 ng) or decreasing the number of target amplification cycles.
Note: Library yield is not indicative of quality.

Find additional tips, troubleshooting help, and resources within our Capillary Electrophoresis Applications Support Center.

Why is my library concentration low with the AgriSeq HTS Library Kit?

Here are possible causes and recommended solutions:
-The sample may have been misquantified. We recommend requantifying the sample DNA using the Qubit 1X dsDNA HS Assay Kit, Qubit dsDNA HS Assay Kit, or the Quant-iT DNA Assay Kit, High Sensitivity and Qubit Fluorometer.
-Residual ethanol from AMPure purification may have inhibited library amplification. We recommend carefully removing all drops of ethanol before library amplification, then centrifuging the plate, if needed. Repeat residual ethanol removal.
-The sample quality may have been low. We recommend adding more DNA or increasing target amplification cycles and ensuring that the input DNA is free of PCR inhibitors. Please contact Technical Support at techsupport@thermofisher.com for details.
-The PCR, digestion, or ligation reactions may have been inefficient. We recommend ensuring proper dispensing and mixing of viscous components at each step.
-The AMPure XP beads may have been overdried. We recommend not drying the AMPure XP beads for more than 1 min.
-The Normalization Beads may not have been washed. We recommend ensuring that the Normalization Beads are washed before use.
-The wrong Normalization Primers may have been used. We recommend using the Normalization Primers that are provided in the AgriSeq HTS Library Kit.
-Residual Normalization Wash may have been present after was. We recommend carefully removing all the Normalization Wash before elution.
Note: Library yield is not indicative of quality.

Find additional tips, troubleshooting help, and resources within our Capillary Electrophoresis Applications Support Center.

What can I use IonCode Barcode Adapters, 1153-1536 for?

When used in combination with other IonCode barcode adapters (Cat. Nos. A31173, A36546, and A44599) and the AgriSeq HTS Library Kit (Cat. Nos. A34143 and A34144), IonCode Barcode Adapters, 1153-1536 enables pooling up to 1,536 amplicon libraries to conduct multiplexed sequencing analysis.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

What is the total hands-on time for the AgriSeq workflow?

The AgriSeq workflow requires less than 4 hours of hands-on time, from sample preparation through data analysis.

Find additional tips, troubleshooting help, and resources within our Capillary Electrophoresis Applications Support Center.