Improved Small RNA Library Preparation Workflow for Next-Generation Sequencing
Poster May 06, 2015
Sabrina Shore, Jordana Henderson, Anton McCaffrey, Gerald Zon, Richard Hogrefe
Next-generation sequencing (NGS) is used to analyze microRNA (miRNA), a class of small non-coding RNAs that are important therapeutic targets and diagnostic markers. Commercially available small RNA sequencing library preparation kits require large inputs (>100 ng) and a laborious gel purification step. Additionally, commercial kits are hindered by adapter dimer formation, where 5' and 3' adapters ligate without an intervening RNA insert. Adapter dimers preferentially amplify during PCR. This is exacerbated at low RNA inputs where adapter dimer formation can greatly diminish usable sequencing reads.
We describe an optimized workflow which suppresses adapter dimers, works with low RNA input and eliminates the need for a gel purification step. Our workflow introduces chemically modified adapters that efficiently form libraries while reducing adapter dimer formation. TriLink’s modified adapter workflow allows RNA inputs as low as 1 ng with less than 1% adapter dimer reads when gel purified. Furthermore, when our workflow is combined with bead-based size selection purification, automation becomes possible.
Psychiatric Risk Gene Cacna1c and Early Life Stress: Potential Gene-Environment interactions?Poster
Early life stress (ELS) is highly associated with development of psychopathology
and mood disorders in adulthood. Genetic studies have identified variation in the gene calcium voltage-gated channel subunit alpha1C (CACNA1C) to increase risk for several psychiatric disorders. This poster assessed the expression of Cacna1c following prepubertal stress.
A new method for generating arrayed RNAi screening tools for any organismPoster
RNA interference (RNAi) using small interfering RNAs (siRNAs) is an important technology for down-regulation of gene expression and a powerful tool to study cellular processes and pathways. Previously, large collections of siRNAs were available only for traditional experimental model systems, such as human and mouse, and predominantly provided as chemically synthesized libraries.READ MORE
A Computational Model of Mood and Future ProspectsPoster
Mood disorders are characterized by changes in reaction to positive events, but studies examining mood in gambling have suggested these changes may not be different from control. Interrogating mood changes with a computational model may give insight into disorders like depression.READ MORE
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