Unique Advantages of the PacBio SMRT Sequencing Platform

Single Molecule Real Time Sequencing can be much more appropriate
for some samples than Illumina or 454 Sequencing.

Here we'll highlight some of the projects we've worked on and pipelines we've generated
for our customers' samples.

Pacific Biosciences Technology

If you're not familiar with Pacific Biosciences' sequencing technology,
take a look at their recent publication or this (soundless) video.

Circular Consensus Sequencing

The ability for the same DNA template to be sequenced multiple times over the course of a sequencing run can overcome the otherwise high error rate (~.13 to .17) of PacBio's technology. By creating circular templates from PCR products or other DNA fragments less than about 2000bp, we can generate thousands of high-quality full-length sequences.

As an example, we have had good success with sequencing diverse PCR products from environmental samples using this technique. From PCR products of using 16S rRNA-specific primers and DNA purified from an array of environmental samples, we have been able to generate thousands of high-quality full length (500-600bp) sequences from each SMRT Cell. We've also developed our own pipeline for rapidly filtering and deriving these sequences from PacBio read data.

Multiple Clustering for Minimally Heterogenous Populations

Sequences of heterogeneous samples with only a few alleles in the population may also be derived with high quality using Multiple Clustering algorithms such as ClustalW. This is useful for PCRs of human DNA with multiple alleles in homogenic sites, or PCRs from populations of just a few organisms such as competition growth assays. We have developed pipelines that can use multiple clustering alignments to derive useful information such as copy number and relative population dynamics from PacBio read data.

Long-Read + Illumina Data for High-Quality Hybrid Assemblies

The average read length for the PacBio platform is increasing at an astounding rate. From some runs over half the reads are already over 5,000 bases long. Because this length spans many of the repetitive elements of most genomes, PacBio data can be used to scaffold contigs generated from Illumina data and De Bruijn Graph-based assemblers to concatenate assembly graphs and produce more closed draft sequences.

Take a look at the different services we offer.