Abstract
Dye-labelled DNA sequencing is one of the most common and robust technique required for molecular biology since 1977 (Sanger, 1977) [1]. I have recently provided the single-step purification method for dye-labeled sequencing products, which is based on the removal of the washing step in EDTA/ethanol precipitation (Fujikura, 2015) [2]. Here I assess and report the accumulated data of the modified method on the larger scale in practice.
Highlights
Dye-labelled DNA sequencing is one of the most common and robust technique required for molecular biology since 1977 (Sanger, 1977) [1]
I have recently provided the single-step purification method for dye-labeled sequencing products, which is based on the removal of the washing step in EDTA/ethanol precipitation (Fujikura, 2015) [2]
The data provide the optimization of methods, including the amount of DNA template, primers, and buffer conditions for rapid purification of dye-labeled sequencing
Summary
The modified EDTA/ethanol method skips the washing step in EDTA/ethanol precipitation for dye-labeled DNA sequencing. The data provide the scheme and characterize the success rate for modified EDTA/ethanol purification method for dye-labeled DNA sequencing. The data provide the optimization of methods, including the amount of DNA template, primers, and buffer conditions for rapid purification of dye-labeled sequencing. The modified method provides the quick and inexpensive purification technique for DNA sequencing. I modified sequencing method and assessed its data quality (Fig. 1). The modified method requires only 10 min, whereas commercial purification kits and standard ethanol precipitation require a longer processing time. DNA sequences of more than 850–900 bp were more stably obtained in a single sequencing reaction with this rapid method without compromising high-quality base calling and read length (916735 bp, n1⁄4168, QV420). The failure rate of modified method was quite low (0.6%; 1/168)
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