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Next Generation Sequencing

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NEXTGEN SEQUENCING

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The following NGS technologies and services are offered:

NGS Platforms

  • Illumina NextSeq 2000:
    This system brings GSC the flexibility for a fast turnaround for smaller projects. With tunable output and high data quality, it provides flexibility for whole-genome, transcriptome, and targeted resequencing. NextSeq 2000 can generate up to 1.8 Billion sequencing reads (Single Reads) on a P4 flow cell. NextSeq Specifications.

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  • Illumina MiSeq: This system is focused on applications such as targeted sequencing, 16S metagenomics, small genome sequencing, targeted gene expression profiling and other amplicon sequencing. Miseq can generate up to 25 million sequencing reads (Single Reads) on a High Output flow cell and sequence 2 x 300 bp read lengths. MiSeq Specifications.

 

  • mRNA-Seq: Stranded and non-stranded, high levels of multiplexing up to 96 or more samples on NovaSeq

    • Standard amounts, Stranded-Seq: 500 ng total RNA, RIN =>8

    • Low Input amounts, Stranded-Seq: 5 ng – 100 ng total RNA

    • Ultra Low Input amounts, Non-Stranded-Seq: 1-1000 cells or 10 pg - 10 ng

  • Total RNA-Seq - Stranded: 5-250 ng

  • Small RNA-Seq: 1 ug, multiplexing up to 48 samples/NextSeq run

  • Partially degraded samples - Stranded and Non-Stranded: LCM, FFPE samples, both stranded and non-stranded, 50 -100 ng

  • Microbial rRNA depletion and RNA-Seq with amounts as low as 1-5 ug of total RNA

  • Whole Genome Sequencing

    • Human / Animal / Plant

    • Microbial

    • As low as 1 ng

  • De novo Sequencing

  • Exome/Targeted capture re-sequencing: Enables high sequencing depths

    • Agilent and Illumina platforms

    • Human, Mouse, Canine and other species

  • Targeted re-sequencing: High levels of multiplexing up to 200 samples / MiSeq run

    • PCR Amplicon sequencing

    • Illumina and Agilent platforms

  • ChIP-Seq

    • Transcription factor analysis

    • Histone modifications

  • DNA Methylation

    • MeDIP- and MBD-Seq

    • MethylC-Seq

    • Agilent SureSelect MethylC-Seq

    • Nucleosome Mapping

      • FAIRE-Seq and DNAse I-Seq

  • 16S amplicon sequencing

  • Whole Genome Metagenomic sequencing

  • Metatranscriptomic analysis

  • DNA/chromatin fragmentation by Covaris

  • DNA / RNA quality analysis: TapeStation assay, Qubit (Picogreen) assays

  • qPCR services

To submit a sample, please complete the appropriate forms and send them to our NGS representative at flsi-illuminaseq-g@vt.edu.

Illumina lists a range of specifications and GSC tries its best to deliver within these specifications. The deliverables also depend on the type (RNA, DNA, low diversity samples etc.) and quality of samples submitted (sub optimal amounts, sample degradation, contamination, whether recommended protocols were used for sample extraction, whether libraries were customer generated etc.). These issues will be discussed before, during and after completion of the project.

 

RNA and DNA QC

  • Includes NanoDrop, Qubit, or TapeStation

  • 2 days to 1 week

  • GSC will send the QC data, and discuss whether the samples can move forward for library preps and sequencing

Library Preparation

  • Includes library preps, QC and quantitation. Please contact the GSC for current turnaround time estimates.

Sequencing

  • Please contact the GSC for current turnaround time estimates.