title: No-wash ethanol precipitation of dye-labeled reaction products improves DNA sequencing reads.
abstract: The advent of DNA sequencing has significantly accelerated molecular biology and clinical genetic testing. Despite recent increases in next-generation sequencing throughput, the most popular platform for DNA sequencing is still the multi-capillary DNA sequencer, which is ideally suited for small-scale sequencing projects and is highly accurate. However, the methods remain time-consuming and laborious. Here, I describe a modified ethylenediaminetetraacetic acid (EDTA) method that skips the washing step in ethanol precipitation. My improvements to standard methods save labor, time, and cost per run and increase the sequence reads by 5 to 10%. This modified method will provide immediate benefits to many researchers.
authors.affiliation: Kobe University Hospital, Chuo-ku, Kobe 650-0017, Japan; Nissei Hospital, Nishi-ku, Osaka 550-0012, Japan. Electronic address: kofujikura@gmail.com.
authors.forename: Kohei
authors.initials: K
authors.lastname: Fujikura
mesh_terms: D011232:Chemical Precipitation; D004396:Coloring Agents; D004492:Edetic Acid; D000431:Ethanol; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D017422:Sequence Analysis, DNA / Q000379:methods*
chemical_list: D004396:Coloring Agents; D000431:Ethanol; D004492:Edetic Acid
keywords: Dye-labeled DNA sequencing; EDTA; Ethanol precipitation; Quality value; Sanger sequencing