SOP62v1_TGD_CUT&RUN and Library prep v1

Abstract

Cleavage Under Targets & Release Using Nuclease (CUT&RUN) is a revolutionary genomic mapping strategy developed by the group of Dr. Steven Henikoff. In CUT&RUN, cells or nuclei are immobilized to a solid support, with pAG-MNase cleaved DNA fragments isolated from solution. The workflow is compatible with next-generation sequencing (NGS) to provide high quality genome-wide profiles of histone post-translational modifications (PTMs) and chromatin-associated proteins (e.g. TFs and chromatin remodelers) Historically, ChIP-seq is the leading approach for genome-wide mapping of histone PTMs and chromatin-associated proteins. ChIP-seq requires large numbers of cells (typically 105 - 106 cells) and deep sequencing of both input chromatin and immunoprecipitated material (typically > 30 million read each) to resolve signal from background. With this innovation, background is dramatically reduces, allowing high resolution genomic mapping for histone PTMs and chromatin-associated proteins using a small number of cells and only 3-8 million sequencing reads per sample. Although it is recommended to start with 500,000 cells, comparable data can be generated using as few as 5,000 cells. Overview of the CUTANA CUT&RUN protocol 1. Immobilize & permeabilize cells (or nuclei) 2. Add antibody to histone PTM or chromatin-interacting protein 3. Add & activate pAG-MNase to cleave target-DNA complex 4. Target-DNA complex diffuses out, collect supernatant 5. Extract DNA & prepare sequencing library 6. Next-generation sequencing and data analysis from CUTANA CUT&RUN kit manual

Comparative assessment of scientific reach and utilization of the International Association of Dental Traumatology 2020 guidelines: An altmetric and citation analysis

“…28 The FCR is a metric specific to Dimensions database, calculated by dividing the number of citations a paper has received by the average number received by documents published in the same year and in the same fields of research (FoR) category. 29 The RCR compares the citation count of an article against a group of articles published through the National Institution of Health (USA) funded articles within the same field. This provides the relative impact of a specific article within its own field of research.…”