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A simple and affordable method for high‐throughput DNA extraction from animal tissues for polymerase chain reaction
Author(s) -
Yue Gen Hua,
Orban Laszlo
Publication year - 2005
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200410411
Subject(s) - throughput , polymerase chain reaction , simple (philosophy) , dna extraction , dna , extraction (chemistry) , computational biology , polymerase , chemistry , chromatography , microbiology and biotechnology , computer science , biology , biochemistry , gene , telecommunications , philosophy , epistemology , wireless
We have developed a very simple and inexpensive method for high‐throughput DNA extraction from animal tissues. The procedure contains three steps (digestion, heating, and centrifugation) and it is compatible with the 96‐well plate format commonly used in polymerase chain reaction (PCR) amplifications. The duration for processing a plate is about 1.5 h; therefore, one researcher can isolate DNA from up to 1000 samples during a single workday. A small piece of tissue ( ca. 10–20 mg) yields enough template for at least 50–70 PCR amplifications, as demonstrated by using the processed samples as templates successfully for long distance PCR, multiplex PCR, and randomly amplified polymorphic DNA (RAPD) assay. The application of our method is expected to facilitate studies that require high‐throughput DNA isolation for PCR amplification, such as genotyping by microsatellites for mapping and genetic diversity studies, as well as mutant screening in zebrafish.