Project description:We investigated the effects of different sample preparation factors on RNA-seq experiments, including RNA concentration, library storage time, and cryopreserved condition, by comparing their sequencing biases, gene expression profiles, and biological function using primary B cell and CD4+ cell blood samples in healthy subjects.
Project description:DNA replication occurs in a defined temporal order known as the replication-timing (RT) program. RT is regulated during development in discrete chromosomal units, coordinated with transcriptional activity and 3D genome organization. Here, we developed a sample preparation method to measure RT genome-wide that does not require fixation.
Project description:A collection of digital automated proteomic sample preparation protocols detail three optimized step-by-step methods to: (A) lyse Gram-negative bacteria and fungal cells; (B) quantify the amount of protein extracted; and (C) normalize the amount of protein and set up tryptic digestion. These protocols have been developed to facilitate rapid, low variance sample preparation of hundreds of samples, be easily implemented on widely-available Beckman-Coulter Biomek automated liquid handlers, and allow flexibility for future protocol development
Project description:Sodium dodecyl sulfate (SDS) is widely used in sample preparation for proteomic research. However, SDS is incompatible with liquid chromatography and electrospray ionization (ESI). SDS depletion is therefore required ahead of liquid chromatography coupled with mass spectrometry (LC-MS) analysis. Most of current SDS removal strategies are time-consuming, laborious and low reproducibility. Here, we describe a method, SDS-cyclodextrin assisted sample preparation (SCASP), by which cyclodextrin can bind to SDS and form cyclodextrins-SDS complexes in solutions, allowing for direct tryptic digestion. We demonstrate that SCASP is a simple, fast and robust SDS-based sample preparation method for proteomics application.