Project description:We selected humann intervertebral disc samples to perform proteomics analysis. There were 1 case of grade I , 1 case of grade II, 3 cases of grade Ⅲ and 3 cases of grade Ⅳ according to Pfirrmann classfication. RNA seqencing analysis and single-cell RNA sequencing were integrated with proteomics data to identify the hub genes for intervertebral disc degeneration using bioinformatic method.
Project description:Total RNA was extracted from human gastric cancer tissues (n=4) and matched adjacent normal tissues (n=4) . RNA samples were analyzed by RNA sequencing based on the manufacturer’s protocols. Briefly, Illumina HiSeq 4000 platform was used to sequence the RNA samples for the subsequent generation of raw data. R package was utilized to select lncRNAs with significantly differential expression based on fold change >2 or <1/2, p value <0.05 between human gastric cancer tissues and matched adjacent normal tissues, and the top 10 upregulated lncRNAs were selected for further study.
Project description:Purposes: To investigate the biological function of tRF in breast cancer by tRF and tiRNA sequencing Methods: Breast cancer tissue samples and matched non-tumor adjacent tissues were obtained from five patients. Small RNA sequencing was performed on Illumina NexSeq instrument Results: If P ≤ 0.05, fold change ≥ 2 as the cut off, there were 3 up-regulated tRFs & tiRNAs and 13 down-regulated tRFs & tiRNAs. Conclusions:There were 3 up-regulated tRFs & tiRNAs and 13 down-regulated tRFs & tiRNAs in breast cancer tissue samples and matching adjacent tissue samples
Project description:Purpose: The goals of this study are using whole-exome and RNA sequencing technologies on 63 cases with TFE3-tRCC to explore the molecular characteristics of TFE3-tRCC and provide potential effective therapeutic strategies Methods: Total RNA was isolated from each sample (63 tumor samples and 14 paired adjacent normal samples) using Qiagen RNeasy formalin-fixed paraffin-embedded (FFPE) Kit (Qiagen, Hilden, Germany). Strand-specific RNA sequencing libraries were generated using the Whole RNA-seq Lib Prep kit for Illumina (ABclonal, China). Final libraries were sequenced at the Novogene Bioinformatics Institute (Beijing, China) on an Illumina Hiseq X10 platform by a 150bp paired-end reads. The raw RNA-sequencing reads were filtered by FastQC, Reads were aligned using STAR v2.7.0f with default parameters to the Ensembl human genome assembly GRCh37. Results: Using an optimized data analysis workflow, we mapped about 30 million sequence reads per sample to the mouse genome (build mm9) and identified 16,014 transcripts in the retinas of WT and Nrl−/− mice with BWA workflow and 34,115 transcripts with TopHat workflow. RNA-seq data confirmed stable expression of 25 known housekeeping genes, and 12 of these were validated with qRT–PCR. RNA-seq data had a linear relationship with qRT–PCR for more than four orders of magnitude and goodness of fit (R2) of 0.8798. Approximately 10% of the transcripts showed differential expression between the WT and Nrl−/− retina, with a fold change ≥1.5 and p value <0.05. Altered expression of 25 genes was confirmed with qRT–PCR, demonstrating the high degree of sensitivity of the RNA-seq method. Hierarchical clustering of differentially expressed genes uncovered several as yet uncharacterized genes that may contribute to retinal function. Data analysis with BWA and TopHat workflows revealed a significant overlap yet provided complementary insights in transcriptome profiling. Conclusions: Our study represents the first detailed analysis of retinal transcriptomes, with biologic replicates, generated by RNA-seq technology. The optimized data analysis workflows reported here should provide a framework for comparative investigations of expression profiles. Our results show that NGS offers a comprehensive and more accurate quantitative and qualitative evaluation of mRNA content within a cell or tissue. We conclude that RNA-seq based transcriptome characterization would expedite genetic network analyses and permit the dissection of complex biologic functions.
