Miniprep for chloroplast and mitochondria enrichment and DNA extraction

Abstract Published protocols for chloroplast and mitochondrial DNA enrichment are based on large samples (more than 5 g) which imply high costs and plenty of sample material. Reduction of tissue sample and reagent quantities is possible and convenient. This work describes an efficient miniprep for chloroplast and mitochondria DNA enrichment and extraction from 500 mg of pericarp samples of Manzano chili fruits (Capsicum pubescens R. & P.). Extraction efficiency from fresh fruit samples was 20 µg DNA g-1 for choloroplasts and 33 µg DNA g-1 for mitochondria. Isolation using the recommended miniprep resulted in costs reduction of 160x for chloroplast and up to 8x for mitochondria. PCR amplification of reference marker genes for chloroplast (rbcL, Rubisco large subunit) and mitochondria (cox1, Cytochrome oxidase) confirmed DNA identity of each extraction.

Bibliographic Details

Main Authors:	Morales-Gutiérrez,Ma. Emma, Gutiérrez-Espinosa,Ma. Alejandra, Santacruz-Varela,Amalio, Ramírez-Ramírez,Iván, Pérez-Grajales,Mario, González-Hernández,Víctor A.
Format:	Digital revista
Language:	English
Published:	Colegio de Postgraduados 2016
Online Access:	http://www.scielo.org.mx/scielo.php?script=sci_arttext&pid=S1405-31952016000800977
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