ABSTRACT: Characterization of flowback and produced waters and long-term accumulated sludges in impoundments, and exploration of an effective pre-treatment method
Project description:The goal of the study was to examine the influence of flowback water exposure on bacterial survival and biocide tolerance. The transcriptome of P. fluorescens was analyzed using RNA-seq to determine the gene rxpression profile changes occuring upon flowback water exposure. The results indicate that that P. fluorescens induces a well-coordinated genetic response that aids in its survival in flowback water as well as imparts enhanced tolerance against typically used slow acting biocides such as gluteraldehyde but increased susceptibity towards sodium hypochlorite. RNA-seq data demonstrated significant induction of genes involved in osmotic stress, energy production and conversion, membrane integrity, protein transport among others.
Project description:The goal of the study was to examine the influence of flowback water exposure on bacterial survival and biocide tolerance. The transcriptome of P. fluorescens was analyzed using RNA-seq to determine the gene rxpression profile changes occuring upon flowback water exposure. The results indicate that that P. fluorescens induces a well-coordinated genetic response that aids in its survival in flowback water as well as imparts enhanced tolerance against typically used slow acting biocides such as gluteraldehyde but increased susceptibity towards sodium hypochlorite. RNA-seq data demonstrated significant induction of genes involved in osmotic stress, energy production and conversion, membrane integrity, protein transport among others. Examination of P. fluorescens transcriptome exposed to flowback water for one hour and compare with PBS exposed cells.
Project description:Hairpin-containing pre-miRNAs are precursors of microRNAs (miRNAs) that play important roles in cellular processes and various human diseases. Pre-miRNAs are produced from longer primary transcripts (pri-miRNAs). The falsity in cellular expression and sequences of pre-miRNAs might cause cellular defects or human diseases. However, the current pre-miRNA quantification methods by qPCR cannot discriminate between pre-miRNAs and their parental pri-miRNAs. In addition, the ligation of the sequencing adapter to 5’-end of pre-miRNAs is inefficient, therefore, pre-miRNA sequencing is highly impractical. Here, we developed a method, called the intramolecular ligation method (iLIME) for pre-miRNA quantification and sequencing. This method utilized T4 RNA ligase 1 to convert hairpin-pre-miRNAs into circularized RNAs that do not naturally exist in cells. The resulting circuliarized RNAs allow us to design unique primers to quantify pre-miRNAs by qPCR specicially, and thus this qPCR can distinguish pre-miRNA from pri-miRNAs. In addition, the iLIME also allows us to sequence pre-miRNAs using next-generation sequencing. The iLIME method offers a simple and effective way to quantify and sequence pre-miRNAs. This will be useful in investigating pre-miRNAs for addressing research questions for medical applications. The iLIME can be potentially extended to other hairpin-containing RNAs, such as tRNAs and snRNAs.