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Our lab is performing single cell RNAseq and mapping the reads back to an existing genome. We are running paired-end sequencing on a NextSeq 500 to analyze differential expression patterns between cell types. I am having difficulty deciding how narrow a range to size select for (using Pippin Prep) after adding adapter sequences. I am aware that tighter size selection reduces bias during bridge amplification, and that the bioinformatics are typically happier with narrower size ranges, but I want to make sure that my size distribution is broad enough to capture a representative sample. What size ranges are people using in their research? Do people feel that selecting transcripts in the 250-300 bp size range would be representative, or should I go tighter/broader? My Covaris sheared libraries currently span 50-1000 bp with the peaks around 300 bp. Any feedback is greatly appreciated!
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The sequencing world is rapidly changing due to declining costs, enhanced accuracies, and the advent of newer, cutting-edge instruments. Equally important to these developments are improvements in sequencing analysis, a process that converts vast amounts of raw data into a comprehensible and meaningful form. This complex task requires expertise and the right analysis tools. In this article, we highlight the progress and innovation in sequencing analysis by reviewing several of the...Yesterday, 07:48 AM -
The field of epigenetics has traditionally concentrated more on DNA and how changes like methylation and phosphorylation of histones impact gene expression and regulation. However, our increased understanding of RNA modifications and their importance in cellular processes has led to a rise in epitranscriptomics research. “Epitranscriptomics brings together the concepts of epigenetics and gene expression,” explained Adrien Leger, PhD, Principal Research Scientist...04-22-2024, 07:01 AM