Abstract
A rapid and economical method of DNA extraction from a red seaweedPorphyra perforata J. Agardh has been developed by the use of lithium chloride. This paper describes the optimization of extraction conditions. Heat treatment of tissues in a solution (0.8 M LiCl, 0.6% Sarkosyl, 10 mM EDTA, 0.2% PVPP, 5% ß-mercaptoethanol, pH 9.0) at 55 °C for 10 min extracts DNA that is of sufficient quality to be used as a template for PCR amplification. Total DNA yield was approximately 30 to 50μg g−1 t of partially dried tissue. Total RNA yield was approximately 400μg g−1 of partially dried tissue. Carbohydrate was contained as approximately 40 to 90 mM (expressed as glucose equivalents) from 1 g tissue, and protein contamination calculated as the O.D. 260/280 ratio was in the range of 1.4 to 1.7. The DNA was characterized by high molecular weight larger than 50 kb.
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Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JD, Smith JA, Struhl K (1987) Current protocols in molecular biology, Greene Publishing Associates and Wiley-Interscience, New York, 3.14.1–3.14.4.
Araki S, Sakurai T, Oohusa T, Sato N (1992) Comparative restriction endonuclease analysis of rhodoplast DNA from different species ofPorphyra (Bangiales, Rhodophyta). Nippon Suisan Gakkaishi. 58:477–480.
Boehringer Mannheim (1990) Reagents for molecular biology (catalog), Boehringer Mannheim Co. Indianapolis, 255 pp.
Bold HC, Wynne MJ (1978) Introduction to the algae. Structure and reproduction, Prentice-Hall Inc. New Jersey, 454 pp.
Brasch DJ, Chang HM, Chuah CT, Melton LD (1981) The galactan sulfate from the edible red algaPorphyra columbina. Carbohydrate Research 97: 113–125.
Evans LV (1963) The use of lithium chloride as a pretreatment to an acetocarmine technique onFucus. Phycologia 2: 187–195.
Fujii T, Naka D, Toyoda N, Seto H (1987) LiCl treatment releases a nickase implicated in genetic transformation ofStreptococcus pneumoniae. J. Bact. 169: 4901–4906.
Hong YK (1993) Cellular differentiation inPorphyra perforata: Aspects of molecular differentiation using the polymerase chain reaction. Ph. D. Dissertation. University of California, Santa Barbara, 135 pp.
Hong YK, Coury DA, Polne-Fuller M, Gibor A (1992) Lithium chloride extraction of DNA from the seaweedPorphyra perforata (Rhodophyta). J. Phycol. 28: 717–720.
John ME (1992) An efficient method for isolation of RNA and DNA from plants containing polyphenolics. Nucl. Acids Res. 20: 2381.
Kendall TL, Byerley DJ, Dean R (1991) Isolation of DNA from blood. Anal. Biochem. 195: 74–76.
Kochert G (1978) Carbohydrate determination by the phenol-sulfuric acid method. In Hellebust JA, Craigie JS (eds), Handbook of Phycological Methods. Vol. II. Physiological and Biochemical Methods. Cambridge University Press, Cambridge, 95–97.
Laird PW, Zijderveld A, Linders K, Rudnicki MA, Jaenisch R, Berns A (1991) Simplified mammalian DNA isolation procedure. Nuc. Acids Res. 19: 4293.
Mayes C, Saunders GW, Tan IH, Druehl LD (1992) DNA extraction methods for kelp (Laminariales) tissue. J. Phycol. 28: 714–716.
Nath K (1990) A rapid DNA isolation procedure from petri dish grown clinical bacterial isolates. Nuc. Acids Res. 18: 6462.
Olsen JL, Stam WT, Bot PVM, van den Hoek C (1987) scDNA-DNA hybridization studies in Pacific and Caribbean isolates ofDictyosphaeria cavernosa (Chlorophyta) indicate a long divergence. Helgol. Meeresunters. 41: 377–83.
Patwary MU, MacKay RM, van der Meer JP (1993) Revealing genetic markers inGelidium vagum (Rhodophyta) through the random amplified polymorphic DNA (RAPD) technique. J. Phycol. 29: 216–222.
Raha S, Merante F, Proteau G, Reed JK (1990) Simultaneous isolation of total cellular RNA and DNA from tissue culture cells using phenol and lithium chloride. Genet. Anal. Tech. Appl. 7: 173–177.
Roell MK, Morse DE (1991) Fractionation of nuclear, chloroplast, and mitochondrial DNA fromPolysiphonia boldii (Rhodophyta) using a rapid and simple method for the simultaneous isolation of RNA and DNA. J. Phycol. 27: 299–305.
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning. 2nd edn. Cold Spring Harbor Laboratory Press, 18.88 pp.
Sato MH, Hisabori T, Yoshida M (1990) The 55-kDa polypeptide released from spinach thylakoid membranes with 1 M LiCl is not the subunit of chloroplast F1. J. Biol. Chem. 265: 13419–13422.
Sogin ML (1990) Application of ribosomal RNA genes for molecular revolution studies. In Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds), PCR protocols: A guide to methods and applications. Academic Press, New York: 307–314.
Su X, Gibor A (1988) A method for RNA isolation from marine macro-algae. Anal. Biochem. 174: 650–657.
Tyobeka EM, Becker RW (1990) Growth and morphological changes induced by lithium chloride treatment of HL-60 cells. Cell Biol. Int. Reports 14: 667–697.
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Hong, YK., Kim, SD., Polne-Fuller, M. et al. DNA extraction conditions fromPorphyra perforata using LiCl. J Appl Phycol 7, 101–107 (1995). https://doi.org/10.1007/BF00693055
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DOI: https://doi.org/10.1007/BF00693055