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Improved nucleic acid extraction protocols for Ganoderma boninense, G. miniatocinctum and G. tornatum.
Nagappan, Jayanthi; Chin, Chiew Foan; Angel, Lee Pei Lee; Cooper, Richard M; May, Sean T; Low, Eng-Ti Leslie.
Afiliación
  • Nagappan J; Malaysian Palm Oil Board, No. 6, Persiaran Institusi, Bandar Baru Bangi, 43000, Kajang, Selangor, Malaysia. jayanthi@mpob.gov.my.
  • Chin CF; School of Biosciences, Faculty of Science, University of Nottingham Malaysia Campus, Jalan Broga, 43500, Semenyih, Selangor, Malaysia. jayanthi@mpob.gov.my.
  • Angel LPL; School of Biosciences, Faculty of Science, University of Nottingham Malaysia Campus, Jalan Broga, 43500, Semenyih, Selangor, Malaysia.
  • Cooper RM; Malaysian Palm Oil Board, No. 6, Persiaran Institusi, Bandar Baru Bangi, 43000, Kajang, Selangor, Malaysia.
  • May ST; Department of Biology & Biochemistry, University of Bath, Bath, BA2 7AY, UK.
  • Low EL; Nottingham Arabidopsis Stock Centre, Division of Plant and Crop Sciences, School of Biosciences, University of Nottingham, Loughborough, UK.
Biotechnol Lett ; 40(11-12): 1541-1550, 2018 Dec.
Article en En | MEDLINE | ID: mdl-30203158
The first and most crucial step of all molecular techniques is to isolate high quality and intact nucleic acids. However, DNA and RNA isolation from fungal samples are usually difficult due to the cell walls that are relatively unsusceptible to lysis and often resistant to traditional extraction procedures. Although there are many extraction protocols for Ganoderma species, different extraction protocols have been applied to different species to obtain high yields of good quality nucleic acids, especially for genome and transcriptome sequencing. Ganoderma species, mainly G. boninense causes the basal stem rot disease, a devastating disease that plagues the oil palm industry. Here, we describe modified DNA extraction protocols for G. boninense, G. miniatocinctum and G. tornatum, and an RNA extraction protocol for G. boninense. The modified salting out DNA extraction protocol is suitable for G. boninense and G. miniatocinctum while the modified high salt and low pH protocol is suitable for G. tornatum. The modified DNA and RNA extraction protocols were able to produce high quality genomic DNA and total RNA of ~ 140 to 160 µg/g and ~ 80 µg/g of mycelia respectively, for Single Molecule Real Time (PacBio Sequel® System) and Illumina sequencing. These protocols will benefit those studying the oil palm pathogens at nucleotide level.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: ADN de Hongos / ARN de Hongos / Ganoderma / Fraccionamiento Químico Idioma: En Revista: Biotechnol Lett Año: 2018 Tipo del documento: Article País de afiliación: Malasia Pais de publicación: Países Bajos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: ADN de Hongos / ARN de Hongos / Ganoderma / Fraccionamiento Químico Idioma: En Revista: Biotechnol Lett Año: 2018 Tipo del documento: Article País de afiliación: Malasia Pais de publicación: Países Bajos