Project description:Chromatin immunoprecipitation coupled with high-throughput sequencing (ChIP-Seq) is a powerful tool to dissect global epigenetic landscapes of cells. However, this method usually consumes millions of cells. Here we develop a robust technique for performing ChIP-Seq using as low as 1,000 cells. This method combines a semi-automatic nanoliter ChIP reaction with a carrier-based sequencing library preparation strategy without pre-amplifying the ChIP product. We used this method to investigate the pattern of trimethylation of histone 3 lysine 4 (H3K4me3) of mouse post-implantation epiblast cells at embryonic day 6.5 (E6.5) and showed that it is more similar to that of mouse Epi-Stem cells (mEpiSCs) than that of mouse Embryonic Stem cells (mESCs). Together with the high similarity between the transcriptomes of EpiSCs and E6.5 epiblast cells, this suggests that mEpiSCs is a reliable in vitro model for post-implantation epiblast cells in vivo. Use 1 million mEpiSCs and 1million mESCs as the positive control, 1000 mEpiSC and 1000 mESCs samples are used to validate the protocol. 1000 mEpiblasts have two biological replicates
Project description:As mechanical stimuli, sound vibrations are known to influence various molecular and physiological processes that can ultimately improve crop performance and yield. Microarray analyses were carried out to investigate the global gene expression changes in MicroTom plants subjected to 1000 Hz sound vibration prior to infection with Pst DC300 to test the effectiveness of sound as a priming agent. Data revealed that 1000 Hz pretreatment is effective in eliciting a transcriptional reprogramming in tomato plants grown under normal conditions, but particularly after Pst DC3000 infection. The protective effect of 1000 Hz pretreatment against Pst DC3000 appeared to be mediated by the upregulation of both unique differentially expressed genes (DEGs) involved in cell wall reinforcement, phenylpropanoid pathway and defensive proteins as well as common DEGs associated with boosting plant basal immunity.
