Sample Requirements

View requirements for the following services:


Microbiome Services

Aliquoting fecal samples

  • Provide fecal sample in collection tub stored at -80C
  • Provide fecal sample in DNA genotek in Omnigene Gut kit stored at room temperature

Nucleic Acid extraction

  • Specify appropriate Qiagen or Promega kit for sample type
  • Provide at least 0.1g of tissue in eppendorf tube stored at -80C
  • Provide at least 250ul of liquid (blood, water or body fluid) stored in -80C
  • Pelleted cells in 1.5ml eppendorf tube stored in -80C

Microbiome Profiling

Library typeLibrary prep kitInput typeRecommended volumeRecommended concentrationRecommended input amountProduct sizeMaximum number of samples pooled per laneRecommended sequencing
16S EMPEMP protocol (V4)DNA2ul5ngs/ul10ngs~390bp192250bp PE MiSeq lane
16S Illumina16S Metagenomic (V3/V4)DNA2.5ul5ngs/ul12.5ngs460bp96300bp PE MiSeq lane
16S Illumina16S Metagenomic (V1-V3)DNA2.55ngs/ul12.5ngs> 600bp96300bp PE MiSeq lane
Illumina DNA seq for bacteriaKappa Hyper PrepDNA25ul20ngs/ul500ngsinsert size ~500bp8150bp PE HiSeq lane
Illumina RNA seq for bacteriaStranded RNA-Seq + Ribo-zero, Epidemiology; human-mouse-rat bacteriatotal RNA10ul50ngs/ul500ngs8150bp PE HiSeq lane

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DNA and RNA Quality CHeck

  • Provide 4uls of sample in eppendorf tubes that are clearly labeled
  • DNA must be stored at -20C
  • RNA must be stored at -80C

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Covaris Shearing

  • Email microbiomesr@duke.edu with the library prep platform/protocol
  • Provide suggested Covaris Shearing protocol, if possible
  • Provide desired fragment length (bp) in online submission form
  • Provide 1.5 - 3ug of genomic DNA or as directed by the library prep protocol in 1.5mL tubes
  • Provide 50ul or 130ul in an eppendorf tube for microTube (for shearing sizes between 2,000 - 5,000 base pairs)
  • Provide 1,000ul in an eppendorf tube for the milliTube

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