Library Preparation

Our libraries are each indexed with unique molecular barcodes. Some library preparations might require additional steps, including shearing, fragmentation, size selection and pooling. The Genomics Core in the NARF at VCU is fully equipped to assist with any size project. Our facility provides full service library construction for multiple applications, including custom projects. Standard protocols in the Genomics Core include:

  • DNA-seq - DNA  (genomic and metagenomic) libraries with different size inserts
  • Amplicon-seq (microbiome analysis uisng 16S rRNA)
  • RNA-seq libraries (polyA+ enriched and strand-specific)
  • RNA-seq libraries (ribosomal depletion and strand-specific)
  • small RNA-seq (microRNA and lncRNAs)
  • Exome capture

For high throughput applications, the Genomics Core uses CaliperTM liquid handling robots to minimize sample handling variation and to provide fast turnaround times. Our next-generation library service follows the same basic steps which include:

  • sample QC (Qubit, BioAnalyzer, FEMTO Pulse or Caliper GX)
  • library preparation (barcoded adaptor ligation)
  • library QC (BioAnalyzer and qPCR)

For libraries prepared by the user, please see "User Prepared Library Guidelines".

 

Input material required

DNA and RNA amounts required for specific applications can be found in the table below. The input DNA and RNA quantities specified below apply if the samples are quantified by a fluorometric method (e.g Qubit, picogreen, ribogreen). If you use the nanodrop spectrophotometer, which habitually over-estimates nucleic acid concentrations, please submit at least five times the requested amount of sample.