Rapid and reliable high-throughput methods of DNA extraction for use in barcoding and molecular systematics of mushrooms

被引:120
作者
Dentinger, Bryn T. M. [1 ,2 ]
Margaritescu, Simona [2 ]
Moncalvo, Jean-Marc [1 ,2 ]
机构
[1] Univ Toronto, Dept Ecol & Evolutionary Biol, Toronto, ON M5S 3B2, Canada
[2] Royal Ontario Museum, Dept Nat Hist, Toronto, ON M5S 2C6, Canada
基金
加拿大自然科学与工程研究理事会;
关键词
FTA; fungi; internal transcribed spacer; proteinase K; rDNA; ribosomal RNA; Whatman; PROTOCOL; SPECIMEN;
D O I
10.1111/j.1755-0998.2009.02825.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We present two methods for DNA extraction from fresh and dried mushrooms that are adaptable to high-throughput sequencing initiatives, such as DNA barcoding. Our results show that these protocols yield similar to 85% sequencing success from recently collected materials. Tests with both recent (< 2 year) and older (> 100 years) specimens reveal that older collections have low success rates and may be an inefficient resource for populating a barcode database. However, our method of extracting DNA from herbarium samples using small amount of tissue is reliable and could be used for important historical specimens. The application of these protocols greatly reduces time, and therefore cost, of generating DNA sequences from mushrooms and other fungi vs. traditional extraction methods. The efficiency of these methods illustrates that standardization and streamlining of sample processing should be shifted from the laboratory to the field.
引用
收藏
页码:628 / 633
页数:6
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