This article has been cited by other articles in PMC. Abstract We observed single DNA molecules at different ethanol concentrations by using fluorescence microscopy. Keywords: DNA structures, ethanol precipitation, phase transitions, single-molecule studies, water nanoclusters. Open in a separate window. Figure 1. Figure 2. Figure 3. Figure 4. Notes Y. References 1. Zeugin J. Crouse J.
Sambrook J. Fregel R. Murrell J. Frank H. Dixit S. Soper A. J, Turner , Soper A. Bowron D. E , 30 , 19—26; [ PubMed ] [ Google Scholar ]. Ueda M. Mikhailenko S. Sergeyev V. Baase W. Sprecher C. Ethanol can also be used to precipitate RNA for much the same reasons, although precipitation of RNA will typically require more ethanol.
Based in San Diego, John Brennan has been writing about science and the environment since Components of Lysis Buffers. Properties of Isopropyl Alcohol. Steps of DNA Transcription. RNA Mutation Vs. DNA Mutation. The Purpose of the Buffer in Electrophoresis. The cost for each method was estimated based on the price of chemicals, enzymes, and disposable items including microfuge tubes and pipette tips consumed for one extraction from a single beetle.
The cost ranges were generated by different prices for various package sizes of the above supplies. The time required to finish one extraction from a single beetle using each method was estimated based on the procedures used in this study, including the time for lysis and 30 min for DNA drying if necessary.
A separate group of beetles including 3 females and 3 males was used for individual total DNA extraction using the SDS method. The supernatant from one single beetle was aliquoted into 15 1. We assumed that the supernatants in the 15 tubes contained an equal amount of DNA. The tubes were treated under a variety of conditions as shown in Table 2. Linear regression and its relevant statistics were used to test the effect of body weight on DNA yield for each extraction method.
The general linear model GLM was applied to test the effects of extraction method and gender on the DNA yield rate and on the absorbance ratio, respectively. GLM was also applied to test precipitation conditions on DNA yield and main effects plot was generated for ethanol volume and temperature evaluated in this study. We thank D. Kondidie UNL for their technical assistance and suggestions. Competing Interests: The authors have declared that no competing interests exist.
Funding: The authors have no support or funding to report. National Center for Biotechnology Information , U. PLoS One. Published online Aug Blair D. Carles Lalueza-Fox, Editor. Author information Article notes Copyright and License information Disclaimer. Received May 30; Accepted Jul 6. Copyright Chen et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
This article has been cited by other articles in PMC. Abstract Background DNA extraction is a routine step in many insect molecular studies. Introduction DNA extraction is a routine step in many biological studies including molecular identification, phylogenetic inference, genetics, and genomics. Results Body weight and DNA yield Although the ranges of body weight overlapped between the females 8.
Table 1 DNA yield rate, absorbance ratio, DNA pellet color, and estimated cost and time used for one beetle extraction by five extraction methods.
Open in a separate window. Figure 1. DNA electrophoresis on 0. PCR amplification of microsatellite loci The core set of six microsatellites, ranging between — bp, were all successfully amplified from the 10 DNA samples extracted by each method data not shown , indicating that the five methods isolated DNA of sufficient quality for PCR application.
Figure 2. Main effects plot of ethanol volume and temperature on DNA yield rate resulted from precipitation. Discussion DNA yield rate, protein contamination, and DNA degradation in the individually-extracted DNA samples from individual beetles varied within each extraction method and across the five methods, although all the methods isolated considerable amounts of DNA with acceptable quality.
DNA extraction protocols For each method, total DNA was individually extracted from 10 beetles, including 5 females and 5 males. Estimation of cost and time required by each method The cost for each method was estimated based on the price of chemicals, enzymes, and disposable items including microfuge tubes and pipette tips consumed for one extraction from a single beetle. Table 2 Treatments of ethanol volume, incubation temperature and time on DNA precipitation.
Data analyses Linear regression and its relevant statistics were used to test the effect of body weight on DNA yield for each extraction method. Acknowledgments We thank D. Footnotes Competing Interests: The authors have declared that no competing interests exist. References 1. Milligan BG. Total DNA isolation. In: Hoelzel AR, editor.
Waldschmidt AM. Braz J Genet. For Study China. Biotechnol Int. Gentra Systems. Rate of turnover of structural variants in the rDNA gene family of Drosophila melanogaster. Mitochondrial DNA in the pine weevil: size, structure and heteroplasmy.
Ribosomal DNA diversity in Apidae. In: Smith DR, editor. Diversity in the Genus Apis.
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