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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 ...
The different stages of the method are lyse, bind, wash, and elute. [1] [2] More specifically, this entails the lysis of target cells to release nucleic acids, selective binding of nucleic acid to a silica membrane, washing away particulates and inhibitors that are not bound to the silica membrane, and elution of the nucleic acid, with the end result being purified nucleic acid in an aqueous ...
Cell isolation is the process of separating individual living cells from a solid block of tissue or cell suspension. While some types of cell naturally exist in a separated form (for example blood cells ), other cell types that are found in solid tissue require specific techniques to separate them into individual cells.
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 ...
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]
RNA extraction is the purification of RNA from biological samples. This procedure is complicated by the ubiquitous presence of ribonuclease enzymes in cells and tissues, which can rapidly degrade RNA. [ 1 ]
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]
18538 Ensembl ENSG00000132646 ENSMUSG00000027342 UniProt P12004 P17918 RefSeq (mRNA) NM_182649 NM_002592 NM_011045 RefSeq (protein) NP_002583 NP_872590 NP_035175 Location (UCSC) Chr 20: 5.11 – 5.13 Mb Chr 2: 132.09 – 132.1 Mb PubMed search Wikidata View/Edit Human View/Edit Mouse Cryo-EM structure of the DNA-bound PolD–PCNA processive complex Proliferating cell nuclear antigen (PCNA) is ...