Improved Small RNA Library Preparation Workflow for Next-Generation Sequencing
Poster Mar 22, 2015
Sabrina Shore, Jordana Henderson, Anton McCaffrey, Gerald Zon, Richard Hogrefe
Next generation sequencing (NGS) is used to analyze microRNAs (miRNAs), 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, which is not amenable to automation. 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 amplification. This is exacerbated at low RNA inputs where adapter dimers can greatly diminish usable sequencing reads. Current methods for adapter dimer suppression are insufficient to allow sequencing of very low input amounts and require a gel purification step to remove adapter dimers. In contrast, we describe an optimized workflow which suppresses adapter dimers, allows for RNA inputs down to 1 ng and eliminates the need for a gel purification step. Our workflow introduces chemically modified adapters that allow efficient library ligation while suppressing 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, the improved workflow using magnetic bead-based size selection allows for automation which was not previously 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.
Novel Role of the Innate Immune DNA Sensor IFI16 (Interferon Gamma Inducible Protein 16) as a Major Epigenetic Modulator During KSHV Infection and Lytic ReactivationPoster
Studies have shown that IFI16 acts as an antiviral restriction factor against a number of DNA viruses, by inhibiting viral replication or transcription through epigenetic modifications. However, till date, no specific epigenetic function of IFI16 has been identified. Here, we have discovered that IFI16 recruits two histone methyltransferases on the KSHV episome leading to altered Histone H3K9 methylation, thus regulating its lifecycle.READ MORE
Correlations among the Toddler Gut Microbiome and Health Status VariablesPoster
Establishment and succession of the human microbiome begins at birth and microbial composition adapts alongside human development and growth. Development of a healthy signature microbial community has significant effects on health.This study followed 18 very low birth weight (VLBW) infants, measured initially in the from Neonatal Intensive Care Unit (NICU), at the age of two to three years old.READ MORE