Visualizing Genetic Variants, Short Targets, and Point Mutations in the Morphological Tissue Context with an RNA <em>In Situ</em> Hybridization Assay

Courtney M. Anderson; Annelies Laeremans; Xiao-Ming Mindy Wang; Xingyong Wu; Bingqing Zhang; Emerald Doolittle; Jeffrey Kim; Na Li; Helly Xiao Yan Pimentel; Emily Park; Xiao-Jun Ma

doi:10.3791/58097

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Visualizing Genetic Variants, Short Targets, and Point Mutations in the Morphological Tissue Context with an RNA In Situ Hybridization Assay

JoVE Journal
Biologia

This content is Free Access.

JoVE Journal Biologia

Visualizing Genetic Variants, Short Targets, and Point Mutations in the Morphological Tissue Context with an RNA In Situ Hybridization Assay

Visualizing Genetic Variants, Short Targets, and Point Mutations in the Morphological Tissue Context with an RNA In Situ Hybridization Assay

DOI:

10.3791/58097-v

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10:57 min

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August 14, 2018

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Courtney M. Anderson, Annelies Laeremans, Xiao-Ming Mindy Wang, Xingyong Wu, Bingqing Zhang, Emerald Doolittle, Jeffrey Kim, Na Li, Helly Xiao Yan Pimentel, Emily Park, Xiao-Jun Ma

¹Advanced Cell Diagnostics, Inc

Capitoli

00:04Titolo
00:41Tissue Preparation
01:30Equipment and Reagent Preparation
03:24Sample Deparaffinization, Dehydration, and Pre-treatment
05:26Probe Hybridization, Signal Amplification, and Detection
07:18Counterstaining, Slide Mounting, and Visualization
08:18Results: Detection of Splice Variants, Short Sequences, and Point Mutation In Situ
10:00Conclusion

Summary

Traduzione automatica

Here, we describe an in situ hybridization assay which enables sensitive and specific detection of sequences as short as 50 nucleotides with single-nucleotide resolution at the single-cell level. The assay, which can be performed manually or automatically, can enable visualization of splice variants, short sequences, and mutations within the tissue context.

Visualizing Genetic Variants, Short Targets, and Point Mutations in the Morphological Tissue Context with an RNA In Situ Hybridization Assay

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