Preparation of Small RNA Libraries for High-Throughput Sequencing | Zendy

Colin D. Malone | Zendy; Julius Brennecke | Zendy; Ben Czech | Zendy; Alexei A. Aravin | Zendy; Gregory J. Han | Zendy

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Preparation of Small RNA Libraries for High-Throughput Sequencing

Author(s) -

Colin D. Malone,

Julius Brennecke,

Ben Czech,

Alexei A. Aravin,

Gregory J. Han

Publication year - 2012

Publication title -

cold spring harbor protocols

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.674

H-Index - 51

eISSN - 1940-3402

pISSN - 1559-6095

DOI - 10.1101/pdb.prot071431

Subject(s) - rna , cloning (programming) , computational biology , dna sequencing , small rna , illumina dye sequencing , chemistry , nucleotide , clone (java method) , dna , biology , microbiology and biotechnology , biochemistry , computer science , gene , programming language

This protocol details the process of small RNA cloning for sequencing on the Illumina/Solexa sequencing platform, but it can be easily modified for use on other next-generation platforms (e.g., SOLiD, 454). This procedure is designed to clone canonical small RNA molecules with 5'-monophosphate and 3'-hydroxyl termini. Modifications, such as the presence of a 2'-O-methyl group, can reduce efficiency, although not sufficiently to negate the utility of the approach. Other termini modifications, such as a 5' triphosphate or a 3' phosphate, can be altered by enzymatic treatment before cloning.

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