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Microfluidic cell sorter sample preparation for genomic assays

Single-cell RNA-Sequencing has led to many novel discoveries such as the detection of rare cell populations, microbial populations, and cancer mutations. The quality of single-cell transcriptomics relies heavily on sample preparation and cell sorting techniques that best preserve RNA quality while r...

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Autores principales: Jagnandan, Nicole, Morachis, Jose
Formato: Online Artículo Texto
Lenguaje:English
Publicado: AIP Publishing LLC 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9188458/
https://www.ncbi.nlm.nih.gov/pubmed/35698630
http://dx.doi.org/10.1063/5.0092358
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author Jagnandan, Nicole
Morachis, Jose
author_facet Jagnandan, Nicole
Morachis, Jose
author_sort Jagnandan, Nicole
collection PubMed
description Single-cell RNA-Sequencing has led to many novel discoveries such as the detection of rare cell populations, microbial populations, and cancer mutations. The quality of single-cell transcriptomics relies heavily on sample preparation and cell sorting techniques that best preserve RNA quality while removing dead cells or debris prior to cDNA generation and library preparation. Magnetic bead cell enrichment is a simple process of cleaning up a sample but can only separate on a single-criterion. Droplet-based cell sorters, on the other hand, allows for higher purity of sorted cells gated on several fluorescent and scatter properties. The downside of traditional droplet-based sorters is their operational complexity, accessibility, and potential stress on cells due to their high-pressure pumps. The WOLF(®) Cell Sorter, and WOLF G2(®), developed by NanoCellect Biomedical, are novel microfluidic-based cell sorters that use gentle sorting technology compatible with several RNA-sequencing platforms. The experiments highlighted here demonstrate how microfluidic sorting can be successfully used to remove debris and unwanted cells prior to genomic sample preparation resulting in more data per cell and improved library complexity.
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spelling pubmed-91884582022-06-12 Microfluidic cell sorter sample preparation for genomic assays Jagnandan, Nicole Morachis, Jose Biomicrofluidics Regular Articles Single-cell RNA-Sequencing has led to many novel discoveries such as the detection of rare cell populations, microbial populations, and cancer mutations. The quality of single-cell transcriptomics relies heavily on sample preparation and cell sorting techniques that best preserve RNA quality while removing dead cells or debris prior to cDNA generation and library preparation. Magnetic bead cell enrichment is a simple process of cleaning up a sample but can only separate on a single-criterion. Droplet-based cell sorters, on the other hand, allows for higher purity of sorted cells gated on several fluorescent and scatter properties. The downside of traditional droplet-based sorters is their operational complexity, accessibility, and potential stress on cells due to their high-pressure pumps. The WOLF(®) Cell Sorter, and WOLF G2(®), developed by NanoCellect Biomedical, are novel microfluidic-based cell sorters that use gentle sorting technology compatible with several RNA-sequencing platforms. The experiments highlighted here demonstrate how microfluidic sorting can be successfully used to remove debris and unwanted cells prior to genomic sample preparation resulting in more data per cell and improved library complexity. AIP Publishing LLC 2022-06-10 /pmc/articles/PMC9188458/ /pubmed/35698630 http://dx.doi.org/10.1063/5.0092358 Text en © 2022 Author(s). https://creativecommons.org/licenses/by/4.0/All article content, except where otherwise noted, is licensed under a Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ).
spellingShingle Regular Articles
Jagnandan, Nicole
Morachis, Jose
Microfluidic cell sorter sample preparation for genomic assays
title Microfluidic cell sorter sample preparation for genomic assays
title_full Microfluidic cell sorter sample preparation for genomic assays
title_fullStr Microfluidic cell sorter sample preparation for genomic assays
title_full_unstemmed Microfluidic cell sorter sample preparation for genomic assays
title_short Microfluidic cell sorter sample preparation for genomic assays
title_sort microfluidic cell sorter sample preparation for genomic assays
topic Regular Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9188458/
https://www.ncbi.nlm.nih.gov/pubmed/35698630
http://dx.doi.org/10.1063/5.0092358
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