Template preparation for rapid PCR in Colletotrichum lindemuthianum

Isolation of DNA for PCR is time-consuming and involves many reagents. The aim of this work was to optimise a rapid and easy PCR methodology without previous DNA isolation. Different strains of the phytopathogenic fungus Colletotrichum lindemuthianum were used. Protoplasts were generated using lytic...

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Bibliographic Details
Published in:	Brazilian journal of microbiology Vol. 34; no. 1; pp. 8 - 12
Main Authors:	Roca, M. Gabriela(Universidade Federal de Lavras Departamento de Biologia), Davide, Lisete C.(Universidade Federal de Lavras Departamento de Biologia), Wheals, Alan E.(University of Bath Department of Biology and Biochemistry)
Format:	Journal Article
Language:	English
Published:	São Paulo Sociedade Brasileira de Microbiologia 01-01-2003 Springer Nature B.V
Subjects:	anthracnose antracnose Colletotrichum lindemuthianum espaços transcritos internamente hygromycin resistance internal transcribed spacer metodolologias de PCR MICROBIOLOGY PCR methodology rapid screening resistência a higromicina triagens rápidas anthracnose triagens rápidas resistência a higromicina hygromycin resistance espaços transcritos internamente rapid screening PCR methodology antracnose internal transcribed spacer metodolologias de PCR
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Summary:	Isolation of DNA for PCR is time-consuming and involves many reagents. The aim of this work was to optimise a rapid and easy PCR methodology without previous DNA isolation. Different strains of the phytopathogenic fungus Colletotrichum lindemuthianum were used. Protoplasts were generated using lytic enzymes under high incubation temperatures using different methodologies to obtain the template. A rapid (10 minute) methodology was successful for smaller amplicons (<750 bp). A extração de DNA para realização de PCR é demorada e envolve vários reagentes. O objetivo deste trabalho foi desenvolver uma metodologia rápida e fácil que permita utilizar a técnica de PCR sem necessidade de extração de DNA. Diferentes isolados de Colletotrichum lindemuthianum foram utilizados. Para obtenção do DNA moldes foram testados protoplastos obtidos através do uso de enzimas líticas, incubados sob altas temperaturas e diferentes metodologias. Uma metodologia rápida (10 minutos) foi desenvolvida para produtos de PCR menores (<750 pb).
Bibliography:	http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822003000100003 10.1590/S1517-83822003000100003 ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23
ISSN:	1517-8382 1678-4405 1678-4405
DOI:	10.1590/S1517-83822003000100003