Next-generation sequencing library preparation for 16S rRNA microbiome analysis after serpin treatment

Juan Maldonado, Jordan R. Yaron, Liqiang Zhang, Alexandra Lucas

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Scopus citations

Abstract

Serine protease inhibitors, serpins, can have profound effects on many systems in the body including immune defense systems. The microbiome, specifically the gut and lung bacterial microbiota, is now known, under some conditions, to alter immune defenses. DNA library preparation for microbiome studies is a procedure that prepares microbial genomic DNA to be sequenced in next-generation sequencing platforms. The construction involves a PCR reaction that will amplify the 16S rRNA gene and will incorporate a specific index and adaptors to the fragments. After confirmation of the product amplification by gel electrophoresis, samples will be later normalized to the same DNA amount of 240 ng. Final concentration of the library is validated by quantitative PCR (qPCR). Here we describe methods to analyze changes in the microbiome after treatment with immune-modulating agents, specifically serpins.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages213-221
Number of pages9
DOIs
StatePublished - 2018

Publication series

NameMethods in Molecular Biology
Volume1826
ISSN (Print)1064-3745

Keywords

  • 16S rRNA gene
  • Library preparation
  • Microbiome
  • Next-generation sequencing
  • Polymerase chain reaction

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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