Learn about SureSelect enrichment libraries |
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The SureSelect enrichment libraries that you can create in eArray let you isolate specific nucleic acid molecules for sequencing. You can create custom libraries of biotinylated RNA "baits" that are complementary to regions of the desired target molecules. The baits capture the desired target molecules by hybridization. Because bait sequences are based on known sequences, these libraries are best used in applications where the underlying sequences are well-characterized, such as in targeted resequencing of genomic DNA that uses next-generation sequencing technology.
eArray has two different SureSelect Enrichment application types: Target Enrichment and RNA Enrichment. Select the application type for your experiments based on the kind of library you want to create. Target Enrichment libraries and RNA Enrichment libraries are described below. Within these two main types of libraries, you can work with several specific types of libraries, which are also described in this help topic.
Target enrichment, also known as genome partitioning, is a method that isolates specific fragments of genomic DNA for sequencing. You use a library of complementary oligonucleotide "baits" to retrieve fragments of interest (target DNA). The target DNA hybridizes well with the baits, but other DNA does not, which forms the basis of a powerful selection method that lets you focus your sequencing efforts.
When you order a SureSelect Target Enrichment library from Agilent, the final product that you receive is a kit that contains a set of biotinylated RNA oligonucleotides. However, as part of the library manufacturing process, Agilent first creates DNA oligonucleotides, and then later transcribes them into RNA. Thus, when you create a library in eArray, you specify bait sequences in terms of DNA bases (A, C, G, T).
SureSelect RNA Enrichment libraries are collections of oligonucleotides that can be used to enrich specific RNA species in a pool of RNAs for sequencing. You use a library of oligonucleotide "baits" that are complementary to the RNAs of interest (target RNAs). The target RNAs hybridize well with the baits, but other RNAs do not, which forms the basis of a powerful selection method that lets you focus your sequencing efforts. The enrichment protocol also preserves the relative abundance of targeted RNAs.
When you order a SureSelect RNA Enrichment library from Agilent, the final product that you receive is a kit that contains a set of biotinylated RNA oligonucleotides. However, as part of the library manufacturing process, Agilent first creates DNA oligonucleotides, and then later transcribes them into RNA. Thus, when you create a SureSelect RNA Enrichment library in eArray, you specify bait sequences in terms of DNA bases (A, C, G, T).
The SureSelect RNA Enrichment application type in eArray supports the Applied Biosystems SOLiD platform and the Illumina sequencing platform. Multiplexing, which lets you sequence multiple samples in the same sequencing reaction, is also supported.
Agilent Catalog libraries – These libraries, available to all registered eArray users, have been designed and tested by Agilent. To see the catalog libraries that are available, do a library search, and select AgilentCatalog as the folder to search.
Custom libraries – These libraries are the ones that you can create in eArray that include the bait content of your choice. You can include baits that already exist in eArray, or those that you upload. You can also create and include new baits based on the tiling of specific transcript sequences or genomic regions of interest.
Normal libraries – "Normal library" is the label that eArray assigns to an Agilent Catalog or custom library if it is not a base library, a combined library, or a custom library set.
Base libraries – A base library is an Agilent Catalog library to which you can add baits for additional transcripts or genomic regions when you create a combined library. Only certain Agilent Catalog libraries can be used as base libraries.
Combined libraries – A combined library contains a selected Agilent Catalog library (the base library) and an additional custom library. This lets you add baits for genomic regions or transcripts that may not be included in the catalog library.
Custom library sets – (SureSelect Target Enrichment application type only) When you design a custom library that contains more baits than can be accommodated in a single library, eArray automatically creates a custom library set. The library set contains as many individual libraries as are needed to accommodate the requested baits.
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