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Home -> Genomics -> NGS Hybridization Capture Kits using Biotinylated RNA Baits (Illumina and other platforms)

NGS Hybridization Capture Kits using Biotinylated RNA Baits (Illumina and other platforms)

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myBaits® target capture kits from Arbor Biosciences provide focused NGS hybridization capture panels for any organism and any project size.

myBaits® is an in-solution NGS library target enrichment system, compatible with many sequencing library types including Illumina®, Ion Torrent®, PacBio and Nanopore generated libraries. A versatile nucleic acid synthesis technology is used to make biotinylated RNA “baits” that are complementary to your sequence targets. Baits and other reagents for NGS target enrichment are supplied with the myBaits kits.

myBaits® kits have been successfully used in thousands of research projects from a wide variety of genome types; animals, plants, and microbes, and DNA sources; fresh, degraded, and environmental. Custom kits tailored for specific target regions are the most popular choice, allowing enrichment of specific SNPs, exons, genes, and other sequence motifs from genomic or metagenomic samples. Arbor Biosciences also offers a variety of predesigned kit options for specific research applications, such as mitochondrial DNA sequencing, ultraconserved element sequencing, whole-genome enrichment from metagenomic samples, and human cancer exome research.


Procedure
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Figure 1. Workflow
(1) HYBRIDIZATION – An NGS library is denatured via heat, and allowed to hybridize to a complex mixture of complementary biotinylated RNA baits over the course of several hours. Adapter-specific blocking oligos prevent random annealing of library molecules at the common adapter sites.
(2) WASHING – After the hybridization is complete, the biotin present on each bait is bound to a streptavidin-coated magnetic bead. Wash steps help remove off-target or poorly-hybridized library molecules.
(3) AMPLIFY – The remaining library molecules that are still bound to their complementary baits are denatured via heat, and amplified using universal library primers. This “enriched” library can now be sequenced.


Typical example of visualized NGS reads

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Figure 2. NGS reads from enriched human gDNA library, aligned to the hg38 genomic reference sequence.
Positions of the four 80nt myBaits probes in this region are indicated by blue bars. Unique read coverage is highest across the baited region, and tapers off upstream and downstream of the baited region. If sequencing further into the known or unknown flanking regions is desired, simply increase the length of your NGS library molecules (compatible with any myBaits kit).

Citations
O.A. Ali et al. (2016) RAD Capture (Rapture): Flexible and Efficient Sequence-Based Genotyping. Genetics 202:389.
M. Giolai et al. (2017) Comparative analysis of targeted long read sequencing approaches for characterization of a plant’s immune receptor repertoire. BMC Genomics 18:564.
J. Starrett et al. (2017) High phylogenetic utility of an ultraconserved element probe set designed for Arachnida. Molecular Ecology Resources 17:812.
J.M. Enk et al. (2016) Mammuthus Population Dynamics in Late Pleistocene North America: Divergence, Phylogeography, and Introgression. Frontiers in Ecology and Evolution.

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