Simultaneous DNA-RNA Extraction from Coastal Sediments and Quantification of 16S rRNA Genes and Transcripts by Real-time PCR

Enrico Tatti; Boyd A. McKew; Corrine Whitby; Cindy J. Smith

doi:10.3791/54067

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ARN-ADN Extraction simultanée de sédiments côtiers et Quantification de 16S ARNr et Transcriptions par PCR en temps réel

Simultaneous DNA-RNA Extraction from Coastal Sediments and Quantification of 16S rRNA Genes and Transcripts by Real-time PCR

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Simultaneous DNA-RNA Extraction from Coastal Sediments and Quantification of 16S rRNA Genes and Transcripts by Real-time PCR

ARN-ADN Extraction simultanée de sédiments côtiers et Quantification de 16S ARNr et Transcriptions par PCR en temps réel

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17,546 Views

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11:37 min

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June 11, 2016

DOI:

10.3791/54067-v

DOI:

10.3791/54067-v

•

11:37 min

June 11, 2016

•

19 Views

Enrico Tatti¹, Boyd A. McKew², Corrine Whitby², Cindy J. Smith¹

¹Microbiology, School of Natural Sciences,National University of Ireland Galway, ²School of Biological Sciences,University of Essex

Transcript

Summary

A protocol to quantify bacterial 16S rRNA genes and transcripts from coastal sediments via real-time PCR is provided. The methodology includes the co-extraction of DNA and RNA; preparation of DNA-free RNA; and 16S rRNA gene and transcript quantification via RT-q-PCR, including standard curve construction.