Single-cell PCR on protargol-impregnated euplotid ciliates: a combined approach of morphological and molecular taxonomy SCIE SCOPUS KCI

Cited 6 time in WEB OF SCIENCE Cited 6 time in Scopus
Title
Single-cell PCR on protargol-impregnated euplotid ciliates: a combined approach of morphological and molecular taxonomy
Author(s)
Kim, Se-Joo; Choi, Joong Ki; Ryu, Seongho; Min, Gi-Sik
Publication Year
2011-09
Abstract
Ciliates are considered one of the most diverse protozoa and play significant roles in ecology. For successful taxonomic study of these microscopic eukaryotes, a staining procedure is necessary, due mainly to intrinsic difficulties in recognizing characteristics from living cells. Although molecular taxonomy has been used to resolve the ambiguities associated with traditional morphology-based taxonomy, extraction of genomic DNA from stained ciliate cells is not available yet. In the present study, we describe a method to extract genomic DNA from a single protargol-impregnated euplotid cell. By using HgCl(2) as a fixative and modulating the exposure time of bleach solution in the protargol impregnation, high-quality genomic DNA can successfully be extracted from a stained single cell with minimal loss of morphological integrity. This technique will contribute to the effectiveness of combined approaches of molecular and morphological taxonomy from single ciliate cells.
ISSN
1976-8354
URI
https://sciwatch.kiost.ac.kr/handle/2020.kiost/3821
DOI
10.1080/19768354.2011.604943
Bibliographic Citation
ANIMAL CELLS AND SYSTEMS, v.15, no.3, pp.251 - 258, 2011
Publisher
TAYLOR & FRANCIS LTD
Subject
RNA GENE-SEQUENCES; SODIUM-HYPOCHLORITE; SPECIES PROBLEM; E-VANNUS; CRASSUS; IDENTIFICATION; DNA; AMPLIFICATION; SPIROTRICHEA; HYPOTRICHIDA
Keywords
euplotids; molecular data; morphology; protargol impregnation; single-cell PCR
Type
Article
Language
English
Document Type
Article
Publisher
TAYLOR & FRANCIS LTD
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