What’s New at the SF
New poster available for download
The Sequencing Facility’s latest informational poster, Innovative Sequencing Resources in the CCR Sequencing Facility, is now available for viewing and downloading through the website. The poster reviews the technological basis and sequencing strengths of the Illumina and PacBio platforms available in our lab, as well as current in-development applications. This is a great resource for PIs seeking a quick overview of the services offered at the SF.
2012 Spring Research Festival
Members of the Sequencing Facility recently participated in a poster session at the NCI-Frederick and Ft. Detrick 2011 Spring Research Festival in Frederick, MD. The festival is an annual opportunity for researchers to share their work with fellow scientists as well as citizens and students from the surrounding Frederick community. The carefully prepared posters developed by our Illumina and Pacbio sequencing teams, describing the procedures and new developments in our pipelines, can be viewed either by clicking the links below or visiting the Protocols and Resources section of our site.
A Pacific Biosciences sequencer has been brought into operation at the Sequencing Facility as part of the Advanced Technology Program’s technology development efforts in early 2011. Pacific Biosciences’ Single Molecule Real Time (SMRT) sequencer, currently in Limited Pre-Release, is the first commercial platform with the ability sequence single template molecules and detect nucleotide incorporations in real time. Each sequencing run takes 15-45 minutes. Up to 48 runs may be loaded at a time for walk away operation. Current protocols in use at the Sequencing Facility include Single Pass sequencing of DNA molecules, maximizing read length (current average read length is ~2000 bp), as well as Circular Consensus sequencing, to maximize the consensus base calling accuracy. Strobe sequencing and modified base detection protocols are currently in development.
Todd Hartley, Jason Mitchell, and Claudia Stewart have developed automation protocols for both 250 bp and 2000 bp library preparations for the Pacific Biosciences RS sequencer. The library preparations allow walk away production of more than one dozen library preparations per day on the BiomekFXP platform. The libraries enable multiple sequencing runs for each library preparation and thus provide complete full time library production for the capacity of the RS sequencer. The quality of the libraries produced by automated and manual methods are similar. The automation team continues to make improvements in the protocol and has delivered basic methods to three external groups for testing. During the next several months the automation group will adapt the methods to the new library preparation protocols and will work on reducing input DNA requirements.
To learn more about the Pacific Biosciences single-molecule DNA sequencer please visit the Pacific Biosciences website or contact Dr. Bao Tran, Director of the Sequencing Facility. A comparative analysis of this and other recently developed next-generation DNA sequencers can be found here.