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Download QR code; Print/export Download as PDF; Printable version; In other projects ... Protocols for Recombinant DNA Isolation, Cloning, and Sequencing
Acid guanidinium thiocyanate-phenol-chloroform extraction (abbreviated AGPC) is a liquid–liquid extraction technique in biochemistry and molecular biology. It is widely used for isolating RNA (as well as DNA and protein in some cases).
It uses a combination of laboratory methods with single-nucleotide transcriptomics in order to compose an atlas of the vascular and perivascular cell types within the brain. The following steps detail the basis of the VINE-Seq protocol: The initial portion of the protocol consists of methodology adapted from splenocyte isolation and sample ...
snRNA-seq uses isolated nuclei instead of the entire cells to profile gene expression. That is to say, scRNA-seq measures both cytoplasmic and nuclear transcripts, while snRNA-seq mainly measures nuclear transcripts (though some transcripts might be attached to the rough endoplasmic reticulum and partially preserved in nuclear preps). [7]
Combining FACS with scRNA-seq has produced optimized protocols such as SORT-seq. [8] A list of studies that utilized SORT-seq can be found here. [9] Moreover, combining microfluidic devices with scRNA-seq has been optimized in 10x Genomics protocols.
A nuclear run-on assay is conducted to identify the genes that are being transcribed at a certain time point. Approximately one million cell nuclei are isolated and incubated with labeled nucleotides, and genes in the process of being transcribed are detected by hybridization of extracted RNA to gene specific probes on a blot. [1]
Like typical next-generation sequencing experiments, single-cell sequencing protocols generally contain the following steps: isolation of a single cell, nucleic acid extraction and amplification, sequencing library preparation, sequencing, and bioinformatic data analysis. It is more challenging to perform single-cell sequencing than sequencing ...
10x Genomics was founded in 2012 by Serge Saxonov, Ben Hindson and Kevin Ness to create advanced testing equipment for use in cellular biology. [3] Prior to starting the company, Saxonov was the founding architect, and director of research and development at 23andMe. [2]