ISSN:
1573-5176
Keywords:
DNA extraction
;
LiCl
;
PCR
;
Porphyra perforata
;
seaweed
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
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.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00693055
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