In situ genomic DNA extraction for PCR analysis of regions of interest in four plant species and one filamentous fungi
Journal Title: Biotecnología Vegetal - Year 2014, Vol 14, Issue 3
Abstract
The extraction methods of genomic DNA are usually laborious and hazardous to human health and the environment by the use of organic solvents (chloroform and phenol). In this work a protocol for in situ extraction of genomic DNA by alkaline lysis is validated. It was used in order to amplify regions of DNA in four species of plants and fungi by polymerase chain reaction (PCR). From plant material of Saccharum officinarum L., Carica papaya L. and Digitalis purpurea L. it was possible to extend different regions of the genome through PCR. Furthermore, it was possible to amplify a fragment of avr-4 gene DNA purified from lyophilized mycelium of Mycosphaerella fijiensis. Additionally, it was possible to amplify the region ap24 transgene inserted into the genome of banana cv. ‘Grande naine’ (Musa AAA).
Authors and Affiliations
Luis E. Rojas , Maritza Reyes , Naivy Pérez-Alonso , María I. Olóriz , Laisyn Posada-Pérez, Bárbara Ocaña , Orelvis Portal , Borys Chong-Pérez , Jorge L. Pérez Pérez
Keywords
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