Rapid Preparation of Filamentous Fungal DNA for PCR Analysis by Ultrasonic Treatment  

作　　者：Gui Yan-ling Han Jie Zhao Jie-hong 

机构地区：[1]College of Pharmacy,Guizhou University of Traditional Chinese Medicine,Guiyang 550021,China

出　　处：《Journal of Northeast Agricultural University(English Edition)》2023年第2期89-96,共8页东北农业大学学报（英文版）

基　　金：Supported by the National Natural Science Foundation of China(81960692);the Science and Technology Support Program of Guizhou Province(2019-2776)。

摘　　要：A simple method to prepare of DNA template suitable for PCR amplification from filamentous fungi will be valuable for improving experimental efficiency.Here,a method was developed which just needed ultrasonic treatment of the mycelium at usual condition,and the produced solution could directly be used as DNA template for internal transcribed spacer(ITS)amplification successfully.The PCR could be improved by additional treatment of 60℃water baths,but was not centrifugation.When the template amount was 0.5-2μL and the ultrasonic time was 7-11 min,there was no distinctly influences on PCR.The method was commonly used for M.purpureus,I.cicadae,Lentinula sp.,Flammul sp.and Dictyophora sp.etc.to detect target sequences of ITS,hygromycin resistance gene(Hyg),CRISPR-associated protein 9(Cas9),Citrinin gene C(CtnC),Citrinin gene D(CtnD),large subunit rRNA gene(NL),and so on.The method could provide a simple,rapid,safe and economic approach to prepare the DNA template for large-scale PCR of the special filamentous fungi materials.

关 键 词：filamentous fungi rapid PCR DNA template preparation ultrasonic treatment 

分 类 号：R197.323[医药卫生—卫生事业管理]