PacBio

The PacBio RS II allows for ultra-long read sequencing, unmatched by other sequencing instruments. The Single Molecule Real-Time (SMRT) technology combines the natural process of DNA replication with real-time observation of DNA synthesis.

The major feature of this sequencer is the ability to produce reads of over on average 15,000 bases. Other leading NGS technologies generate relative short reads (150 basepairs). The PacBio RS II is therefore the method of choice when characterizing structural variations, repeat regions, pseudogenes and structural variation characterization. Long range sequencing is also the method of choice for de novo sequencing of small genomes, such as micro-organisms. 

Advantages:

  • Ultra long reads of up to 80 Kb
  • Uniform coverage
  • Easy filtering due to random errors
  • No GC bias
  • Unique in detecting base modifications

Technology Background
Zero-Mode Waveguides (ZMWs) allow light to illuminate only the bottom of a well that holds a DNA polymerase/template complex. SMRT cells contain thousands of ZMWs in which the waveguides are monitored simultaneously in real time. The reads are read by the Circular Consensus Sequencing (CCS) technique, in which the same DNA loop is read multiple times.

Why choose PacBio at GenomeScan:

  • Closure of incomplete genome assemblies, especially for polyploids such as vegetables
  • Reduced number of contigs and scaffolding due to easy mapping of long reads
  • Accurate characterization of large structural variations
  • Best option to sequence repeat regions and pseudogenes
  • Balanced genome coverage with minimal GC bias

Custom solutions
We offer custom solutions such as the combination of HiSeq and PacBio sequencing. This option is ideal for de novo assemblies where the long PacBio reads serve as a reference to align the smaller but more accurate HiSeq reads to. We successfully applied this method for notoriously difficult genomes such as the polyploid genome of the onion.