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DNA EXTRACTION METHODS FOR KELP (LAMINARIALES) TISSUE 1
Author(s) -
Mayes Charlene,
Saunders Gary W.,
Tan Ian H.,
Druehl Louis D.
Publication year - 1992
Publication title -
journal of phycology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.85
H-Index - 127
eISSN - 1529-8817
pISSN - 0022-3646
DOI - 10.1111/j.0022-3646.1992.00712.x
Subject(s) - gametophyte , biology , sporophyte , kelp , dna extraction , dna , botany , restriction enzyme , extraction (chemistry) , laminaria , polymerase chain reaction , algae , biochemistry , chromatography , gene , pollen , chemistry
Newly adapted extraction methods for harvesting total cellular DNA from kelp (Laminariales; Phaeophyta) exploit the life‐history stages of these heteromorphic algae. Earlier techniques, developed primarily for chloroplast DNA extraction, were time consuming and labor intensive and required large quantities of fresh sporophyte tissue. In contrast, the new methods expedite DNA extraction by employing dried sporophyte laminae or fresh gametophyte filaments and meiospores. The current methods require less tissue and procedural manipulation, reducing the time and labor involved in DNA extraction. Additionally, the current methods were successfully employed to extract DNA from herbarium specimens up to 22 years old. Total cellular DNA yields were 12.8 and 26.4 μg:g −1 from dried laminae,: 1.9 and 11 μg from small volumes of concentrated gametophyte filaments (25 μL), and 43 to 54.5 μg from pellets of isolated meiospores (50 μL). Following purification on Sepharose columns, DNA from all three sources was sufficiently pure for molecular applications, including restriction endonuclease digestion, Southern blot‐hybridizations, and amplification via the polymerase chain reaction.