Efficacy of Using a Combination Microplate Washer for Vacuum-Based DNA Sequencing Reaction Cleanup
Poster Feb 16, 2012
Wendy Goodrich, Jason Greene, Mary Louise Shane
Over the past 10-12 years the evolution of market leading dye terminator methods and automated capillary electrophoresis instrumentation has largely standardized the procedure for DNA sequencing, quickly making it more accessible, less resource intensive, and easier to perform at many different throughput levels. A critical component of this genomic workflow is the sequencing cleanup procedure, where contaminating artifacts of the sequencing reaction are removed prior to capillary electrophoresis. There are currently a number of viable DNA sequencing cleanup methods available using either filtration, precipitation, or sequestering. Each method has its own costs and benefits and is a proven way of purifying reaction samples. In collaboration with a comprehensive DNA Analysis Core Facility that utilizes state-of-the-art sequencing chemistires and technology, a microplate washer fitted with an integrated vacuum filtration module was used to perform DNA sequencing reaction cleanup. Results were substantiated against a gel filtration method currently used by the collaborator. Evidence provided by this demonstration support the efficacy of the microplate washer demonstrated here to contribute to the genomic workflow typical of many molecular biology laboratories and core facilities.
When there is a need to quickly analyze samples using a number of different PCR assays, it is likely that optimal conditions for each assay will not be the same. First, different assays often will require different annealing temperatures for their primers. In addition, amplicons may be designed to be of different lengths and therefore require varying durations of the extension step.READ MORE