Massive Parallel Sequencing

 

iontorrent illumina roche454

VIB Nucleomics Core offers Illumina Next Generation Sequencing platforms, with great flexibility in running both small and large scale projects within a reasonable time.

 

Most appropriate applications

  • genome sequencing (MiSeq suited for small genomes only)
  • exome sequencing (not MiSeq)
  • digital gene expression (MiSeq suited for small transcriptomes only)
  • transcriptomics (small transcriptomes on MiSeq)
  • small RNA sequencing
  • targeted resequencing
  • MiSeq: QC for HiSeq libraries

Steps in each sequencing project, irrespective of the platform or application:

  1. library preparation
  2. bridge amplification
  3. the sequencing itself

 

Overview of the available sequencing equipment and their capacity

 

Instrument Time Read length Lanes Max #reads Max cap.
Illumina NextSeq 30 h 1*75 – 2*150 bp 4 technical lanes 400*106 120 Gbp
Illumina MiSeq 5.5 – 55 h 1*50 – 2*300 bp 1 25*106 14 Gbp

 

 

Sequencing library preparation

The sequencing library preparation is highly dependent on the application and often multiple vendors are offering slightly different library preparation kits. VIB Nucleomics Core is constantly implementing these library preparation protocols, preferentially on an automated platform. Especially with targeted resequencing, many options are available to enrich for sequences of interest to the researcher. PCR based approaches are very well suited for the smaller target regions (< 200 kbp).

Companies offering PCR based enrichment solutions:

 

Sequencing capacity

The sequencing capacity of the MiSeq fits very well with required coverage of smaller-sized target regions. Larger target regions, such as whole exomes, benefit more from hybridization based enrichment. Those experiments will need the sequencing capacity from the Illumina NextSeq.

Companies offering hybridization based enrichment:

 

Outsourcing

Apart from running sequencing services in-house on 454, IonTorrent or Illumina, VIB Nucleomics Core, through its contacts with other service providers, also can offer project management involving outsourcing to established companies like Complete Genomics, BGI or for evaluation to technology providers building novel platforms (e.g. nanopore technology or Pacific Biosciences).

 

Experimental design, sample requirements, and data analysis expectations

Clearly, sequencing is currently a rapidly evolving field with already now a wealth of different applications and options to the researcher.   Therefore we highly recommend to contact us and to consult us on experimental design, sample requirements, and data analysis expectations.

Sample submission

 

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