Friday, April 1, 2016

How does SMALL RNA SEQUENCING works?


Non-coding small RNAs (ncRNA, smallRNA) and microRNAs (miRNA) are key regulators of gene expression, heterochromatin formation and nucleus organisation. Their action is dependent on tissue and environment and, through various mechanisms, they have a major influence on cancer and numerous diseases.
Several platforms within the France Génomique network will accept requests for small RNA studies or microRNA studies (Pasteur, Strasbourg, Montpellier, Nice, Paris centre, CEA/IG-CNG).
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Starting with total RNA, the small RNAs are purified on acrylamide gel and used to prepare sequencing libraries. In this step, a sequence specific adapter is added to both ends of the RNA molecule, RNA is converted into double-stranded cDNA, and amplification is performed. The libraries are then quality controlled on a Bioanalyser (Agilent) and quantified on a QuBit (Invitrogen). The resulting sequences will be of native RNA (directional RNA-Seq).