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.
Despite the developments in conventional PCR, the complexity of multiplex Real Time PCR is still limited due to the lack of sufficient detection channels. To achieve high-end multiplexing capacity on standard Real Time PCR machines, Anapa Biotech has developed the MeltPlex® technology (see box on right).READ MORE
Genome-wide association studies (GWAS) have identified more than 100 genetic loci associated with type 2 diabetes. The majority of these are located in the intergenic or intragenic regions suggesting that the implicated variants may alter chromatin conformation. This, in turn, is likely to influence the expression of nearby or more remotely located genes to alter beta cell function. At present, however, detailed molecular and functional analyses are still lacking for most of these variants. We recently analysed one of these loci and mapped five causal variants in an islet-specific enhancer cluster within the STARD10 gene locus. Here, we aimed to understand how these causal variants influence b-cell function by alteration of the chromatin structure of enhancer clusterREAD MORE