A simple and affordable method for high-throughput DNA extraction from animal tissues for polymerase chain reaction.
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.
Yue
Gen Hua
GH
Reproductive Genomics Group, Temasek Life Sciences Laboratory, 1 Research Link, National University of Singapore, Singapore 117604.
Orban
Laszlo
L
D000818:Animals; D002347:Carps; D005591:Chemical Fractionation / Q000379:methods*; D004247:DNA / Q000302:isolation & purification*; D005399:Fishes; D018895:Microsatellite Repeats; D009132:Muscles / Q000737:chemistry; D016133:Polymerase Chain Reaction / Q000379:methods*; D019105:Random Amplified Polymorphic DNA Technique; D013552:Swine; D015027:Zebrafish / Q000196:embryology
D004247:DNA