Project description:This experiment was designed to improve the proportion of miRNA mapping reads from sRNA library sequencing, by reducing the amount of particularly abundant rRNA fragments.
Project description:This experiment was designed to improve the percentage of miRNA mapping reads from sRNA library sequencing, by reducing the amount of one particularly abundant rRNA 30mer sequence.
Project description:We report the different expression level of in vivo transcriptome and DMEM cultured bacteria E.coli O157:H7 samples. After obtaining RNA from mice colon and LB cultured free living cells, RNA samples were purified using an RNeasy Mini Kit (Qiagen) and bacterial rRNAs were depleted using the Ribo-Zero rRNA Removal Kit (Epicentre; #RZNB1056). Library construction of RNA samples was carried out using a NEBNext R UltraTM Directional RNA Library Prep Kit for Illumina R (NEB, USA), following the manufacturer’s recommendations. After cluster generation, library preparations were sequenced on an Illumina Hiseq platform to generate paired-end reads.We find that a novel sRNA s77 significantly upregulated compared in vivo samples and LB-cultured sample. This study provides a novel sRNA for further identifications.
Project description:A spectral library was built for Drosophila melanogaster. The spectral library allows reproducible quantification for thousands of peptides per SWATH-MS analysis.
Proteins from Drosophila melanogaster embryo, adult flies were digested with trypsin using in-gel digestion and the peptides were fractionated by high-pH reverse phase chromatography. HRM peptides were spiked into the peptides mixture and each fraction was injected on a Sciex TripleTOF 6600 mass spectrometer fitted with microflow set-up.
The resulting .wiff files were analysed using MaxQuant and Spectronaut.
Project description:Small RNAs (sRNAs) play important roles in plants encountering stress environments. However, limited research has been conducted on the sRNAs involved in plant wound responses. To identify potential roles for the wounding-related sRNAs, sRNA deep sequencing was used. After leaves were wounded for 0.5 hour, total RNAs from unwounded and wounded leaves were isolated for sRNA library construction. The Illumina platform was used to sequence sRNA libraries. About 12 million sequence reads were obtained for each sample.
Project description:To understand the role of message RNA (mRNAs, lncRNAs and circRNAs) in regulation of intramuscular fat deposition, the expression profiles of longissimus dorsi muscle from six Laiwu pigs were sequenced based on rRNA-depleted library construction. We then performed gene expression profiling analysis using data obtained from RNA-seq.
Project description:Multiple companies produce RecJ exonuclease and RNA dependent DNA polymerase for making cDNA. Our standard protocol was developed based on the enzymes from Epicenter (epicenter was recently bought by the Illumina company). As an alternative for the enzymes from EPicenter, we tested the RecJ exonuclease from NEB and Superperscript II from Invitrogen in sRNA library construction using HD adapters [Sorefan et al 2012, Xu et al 2015].
