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Guided Protocol for Fecal Microbial Characterization by 16S rRNA-Amplicon Sequencing
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Guided Protocol for Fecal Microbial Characterization by 16S rRNA-Amplicon Sequencing

Guided Protocol for Fecal Microbial Characterization by 16S rRNA-Amplicon Sequencing

DOI:
10.3791/56845-v

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08:05 min

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March 19, 2018

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, , , , , , , ,
1Sheba Medical Center, 2Children’s Hospital of Pittsburgh of UPMC, 3Ariel University, 4Cincinnati Children’s Hospital Medical Center

Chapters

  • 00:04Title
  • 00:59Sample Handling and DNA Extraction
  • 02:12PCR and Library Preparation
  • 05:30Results: 16S RNA-Seq of Prepared Libraries
  • 07:19Conclusion

Summary

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This manuscript describes a detailed standardized protocol of high-throughput 16S rRNA-amplicon sequencing. The protocol introduces an integrated, uniformed, feasible, and inexpensive protocol starting from fecal sample collection through data analyses. This protocol enables analysis of large numbers of samples with rigorous standards and several controls.

Tags

16S RRNA-Amplicon SequencingFecal MicrobiomeMicrobial CompositionAerobic And Anaerobic CommunitiesHigh-throughputFecal Sample ProcessingDNA ExtractionPCRPrimer Design96-well Plate SetupTriplicate ReactionsPCR Master Mix

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