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Optimized single-cell RNA sequencing protocol to study early genome activation in mammalian preimplantation development. | LitMetric

Optimized single-cell RNA sequencing protocol to study early genome activation in mammalian preimplantation development.

STAR Protoc

Department of Biosciences and Nutrition, Karolinska Institutet, 14183 Huddinge, Sweden; Folkhälsan Research Center, 00290 Helsinki, Finland; Stem Cells and Metabolism Research Program, University of Helsinki, 00290 Helsinki, Finland. Electronic address:

Published: September 2023

AI Article Synopsis

Article Abstract

Here, we present a modification of single-cell tagged reverse transcription protocol to study gene expression on a single-cell level or with limited RNA input. We describe different enzymes for reverse transcription and cDNA amplification, modified lysis buffer, and additional clean-up steps before cDNA amplification. We also detail an optimized single-cell RNA sequencing method for handpicked single cells, or tens to hundreds of cells, as input material to study mammalian preimplantation development. For complete details on the use and execution of this protocol, please refer to Ezer et al..

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10277609PMC
http://dx.doi.org/10.1016/j.xpro.2023.102357DOI Listing

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