Improved Flow Cytometric Analysis of the Budding Yeast Cell Cycle

被引：195

作者：

Haase, Steven B. ^{[1
]}

Reed, Steven I. ^{[1
]}

机构：

[1] Scripps Res Inst, Dept Mol Biol, La Jolla, CA 92037 USA

来源：

CELL CYCLE | 2002年 / 1卷 / 02期

关键词：

Flow cytometry; Cell cycle; Saccharomyces cerevisiae; SYTOX Green; Propidium iodide;

D O I：

10.4161/cc.1.2.114

中图分类号：

Q2 [细胞生物学];

学科分类号：

071009 ; 090102 ;

摘要：

The budding yeast, Saccharomyces cerevisiae has been a remarkably useful model system for the study of eukaryotic cell cycle regulation. Flow cytometric analysis of DNA content in budding yeast has become a standard tool for the analysis of cell cycle progression. However, popular protocols utilizing the DNA binding dye, propidium iodide, suffer from a number of drawbacks that confound accurate analysis by flow cytometry. Here we show the utility of the DNA binding dye, SYTOX Green, in the cell cycle analysis of yeast. Samples analyzed using SYTOX Green exhibited better coefficients of variation, improved linearity between DNA content and fluorescence, and decreased peak drift associated with changes in dye concentration, growth conditions or cell size.

引用

页码：132 / 136

页数：5

共 10 条

[1] Mec1p regulates Pds1p levels in S phase: complex coordination of DNA replication and mitosis [J].

Clarke, DJ ;

Segal, M ;

Jensen, S ;

Reed, SI .

NATURE CELL BIOLOGY, 2001, 3 (07) :619-627

[2] CHARACTERIZATION OF 4 B-TYPE CYCLIN GENES OF THE BUDDING YEAST SACCHAROMYCES-CEREVISIAE [J].

FITCH, I ;

DAHMANN, C ;

SURANA, U ;

AMON, A ;

NASMYTH, K ;

GOETSCH, L ;

BYERS, B ;

FUTCHER, B .

MOLECULAR BIOLOGY OF THE CELL, 1992, 3 (07) :805-818

[3]

Foss EJ, 2001, GENETICS, V157, P567

[4]

Haase SB, 1997, METHOD ENZYMOL, V283, P322

[5]

HARTWELL LH, 1985, GENETICS, V110, P381

[6]

Mathias N, 1996, MOL CELL BIOL, V16, P6634

[7] Cyclin-dependent kinases prevent DNA re-replication through multiple mechanisms [J].

Nguyen, VQ ;

Co, C ;

Li, JJ .

NATURE, 2001, 411 (6841) :1068-1073

[8]

Rhind N, 1998, GENETICS, V149, P1729

[9] CYCLIN-B HOMOLOGS IN SACCHAROMYCES-CEREVISIAE FUNCTION IN S-PHASE AND IN G(2) [J].

RICHARDSON, H ;

LEW, DJ ;

HENZE, M ;

SUGIMOTO, K ;

REED, SI .

GENES & DEVELOPMENT, 1992, 6 (11) :2021-2034

[10]

Sousa M, 2001, CELL CYCLE ANAL YEAS, V1

← 1 →