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The purpose of using RNA FISH is to detect target mRNA transcripts in cells, tissue sections, or even whole-mounts. [10] The process is done in 3 main procedures: tissue preparation (pre-hybridization), hybridization, and washing (post-hybridization). The tissue preparation starts by collecting the appropriate tissue sections to perform RNA FISH.
RNA-Seq (named as an abbreviation of RNA sequencing) is a technique that uses next-generation sequencing to reveal the presence and quantity of RNA molecules in a biological sample, providing a snapshot of gene expression in the sample, also known as transcriptome.
FISSEQ combines the spatial context of RNA-FISH and the global transcriptome profiling of RNA-seq. [1] FISSEQ preserves the tissue allowing single molecule in situ RNA localization. The foundation of the method is a novel nucleic acid sequencing library construction method that stably cross-links cDNA amplicons within biological samples. [2]
Megaselia abdita, scuttle fly (transcriptome 2013 [397]) Family Psychodidae (drain flies) Clogmia albipunctata, moth midge (transcriptome 2013 [397]) Family Sarcophagidae (flesh flies) Sarcophaga Bullata, Flesh fly (2019 [398]) Family Syrphidae (hoverflies) Episyrphus balteatus, hoverfly (transcriptome 2011 [399]) Order Hemiptera
The word "transcriptome" was first used in the 1990s. [19] [20] In 1995, one of the earliest sequencing-based transcriptomic methods was developed, serial analysis of gene expression (SAGE), which worked by Sanger sequencing of concatenated random transcript fragments. [21] Transcripts were quantified by matching the fragments to known genes.
Single-cell RNA sequencing (scRNA-seq) is a recently developed technique that allows the analysis of the transcriptome of single cells, including bacteria. [25] With single-cell transcriptomics, subpopulations of cell types that constitute the tissue of interest are also taken into consideration. [ 26 ]
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