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Literature DB >> 34179277

Terminal Deoxynucleotidyl Transferase Mediated Production of Labeled Probes for Single-molecule FISH or RNA Capture.

Imre Gaspar¹, Frank Wippich¹, Anne Ephrussi¹.

Abstract

Arrays of short, singly-labeled ssDNA oligonucleotides enable in situ hybridization with single molecule sensitivity and efficient transcript specific RNA capture. Here, we describe a simple, enzymatic protocol that can be carried out using basic laboratory equipment to convert arrays of PCR oligos into smFISH and RAP probesets in a quantitative, cost-efficient and flexible way.

Entities: Chemical

Keywords: Labelled terminator nucleotide; Probe production; RNA affinity purification (RAP); RNA capture; Single molecule FISH (smFISH); Terminal deoxynucleotidyl transferase

Year: 2018 PMID： 34179277 PMCID： PMC8203885 DOI： 10.21769/BioProtoc.2750

Source DB: PubMed Journal: Bio Protoc ISSN： 2331-8325

10 in total

1 in total

1. Single-molecule Fluorescence in situ Hybridization (smFISH) for RNA Detection in Adherent Animal Cells.

Authors: Gal Haimovich; Jeffrey E Gerst
Journal: Bio Protoc Date: 2018-11-05

1 in total

Terminal Deoxynucleotidyl Transferase Mediated Production of Labeled Probes for Single-molecule FISH or RNA Capture.

1. Visualization of single RNA transcripts in situ.

Review 2. DeconvolutionLab2: An open-source software for deconvolution microscopy.

3. The Stress Granule Transcriptome Reveals Principles of mRNA Accumulation in Stress Granules.

4. Ex vivo Ooplasmic Extract from Developing Drosophila Oocytes for Quantitative TIRF Microscopy Analysis.

5. Imaging individual mRNA molecules using multiple singly labeled probes.

6. Independent and coordinate trafficking of single Drosophila germ plasm mRNAs.

7. Systematic imaging reveals features and changing localization of mRNAs in Drosophila development.

Review 8. Strength in numbers: quantitative single-molecule RNA detection assays.

9. Enzymatic production of single-molecule FISH and RNA capture probes.

10. An RNA-binding atypical tropomyosin recruits kinesin-1 dynamically to oskar mRNPs.

1. Single-molecule Fluorescence in situ Hybridization (smFISH) for RNA Detection in Adherent Animal Cells.