Isolation of Intact Chloroplast for Sequencing Plastid Genomes of Five Festuca Species.

ABSTRACT: Isolation of good quality chloroplast DNA (cpDNA) is a challenge in different plant species, although several methods for isolation are known. Attempts were undertaken to isolate cpDNA from Festuca grass species by using available standard protocols; however, they failed due to difficulties separating intact chloroplasts from the polysaccharides, oleoresin, and contaminated nuclear DNA that are present in the crude homogenate. In this study, we present a quick and inexpensive protocol for isolating intact chloroplasts from seven grass varieties/accessions of five Festuca species using a single layer of 30% Percoll solution. This protocol was successful in isolating high quality cpDNA with the least amount of contamination of other DNA. We performed Illumina MiSeq paired-end sequencing (2 ×

SUBMITTER: Islam MS 

PROVIDER: S-EPMC6963596 | biostudies-literature | 2019 Dec

REPOSITORIES: biostudies-literature