title: Analysis of the DNA sequencing quality and efficiency of the Apollo100 robotic microcycler in a core facility setting.
abstract: Sanger, or dideoxynucleotide sequencing, is an important tool for biomolecular research. An important trend in DNA sequencing is to find new and innovative ways to provide high-quality, reliable sequences in a more efficient manner, using automated capillary electrophoresis. The Apollo100 combines Sanger cycle sequencing and solid-phase reversible immobilization for product purification in a single instrument with robotic liquid handling and microfluidic (Microscale On-chip Valve) chips that have onboard thermal cycling and pneumatic mixing. Experiments were performed to determine how the DNA sequencing results from the Apollo100 compared with conventional, manual methods used in a core facility setting. Through rigorous experimentation of multiple baseline runs and a dilution series of template concentration, the Apollo100 generated sequencing that exceeded 900 bases with a quality score of 20 or above. When comparing actual client samples of amplicons, plasmids, and cosmids, Apollo100 sequencing results did not differ significantly from those reactions prepared manually. In addition, bacterial genomic DNA was sequenced successfully, directly with the Apollo100, although results were of lower quality than the standard manual method. As a result of the microscale capabilities, the Apollo100 offers valuable savings with respect to the quantity of reagents consumed compared with current manual sequencing methods, thereby continuing the demand for smaller template and reagent requirements. In conclusion, the Apollo100 can generate high-quality DNA sequences for common templates equivalent to those produced using manual sequencing methods and increases efficiency through reduced labor and reagents.
authors: Plant-Microbe Genomics Facility, The Ohio State University, Columbus, USA.
authors: M E
authors: ME
authors: Logsdon
authors: M C
authors: MC
authors: Trounstine
authors: M R
authors: MR
authors: Zianni
mesh_terms: D057205:Automation, Laboratory / Q000295:instrumentation*; D001407:Bacillus / Q000235:genetics; D001483:Base Sequence; D019075:Electrophoresis, Capillary; D016680:Genome, Bacterial; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA / Q000295:instrumentation*