(http://www.vermont211.org/ or http://www.vermontredcross.org/). It is time to heal, repair, cleanup.
Scientifically speaking of cleanup, segue to my blog posting this week…DNA sequencing cleanup. Although Next Generation Sequencing garners a lot of attention, traditional sequencing based on the Sanger method is still an indispensable part of contemporary molecular biology. Over the past 10-12 years the evolution of market leading dye terminator methods and automated capillary electrophoresis instrumentation has largely standardized traditional sequencing. A critical component of this workflow is the cleanup procedure, where contaminating artifacts from the sequencing reaction are removed prior to capillary electrophoresis. There are currently a number of viable sequencing cleanup methods available using either filtration, precipitation, or sequestering as a process of choice.
BioTek is making a foray into the cleanup procedures of the genomic workflow, demonstrating first a filtration method for sequencing cleanup. The BioTek ELx405 vacuum filtration module was utilized with a Millipore membrane based size-exclusion kit to compare performance to a matrix gel filtration method used by the Vermont Cancer Center DNA Analysis Facility affiliated with the University of Vermont and Fletcher Allen Healthcare. We were fortunate enough to work with very experienced staff using state of the art technology for this application. A poster presentation at European Lab Automation in June 2011 highlighted the results of this collaboration. An Application Note will also be published soon offering additional information for optimizing vacuum performance among other lessons learned. A Quality Matrix and chromatogram of comparative results are shown below.
The promising outcomes of this collaboration have led to additional research investigating the potential viability of our ELx405 and EL406 for performing magnetic bead based sequencing cleanup, a nice companion piece to the filtration methods already demonstrated. To round out this niche market application analysis stay appraised for updates on a potential mag bead pcr cleanup demonstration.