Flow cytometry assay for the detection of single-copy DNA in human lymphocytes

被引：2
|
作者：
Uno, Naoki [1 ]
Kaku, Norihito [1 ]
Morinaga, Yoshitomo [1 ]
Hasegawa, Hiroo [1 ]
Yanagihara, Katsunori [1 ]
机构：
[1] Nagasaki Univ, Dept Lab Med, Grad Sch Biomed Sci, Nagasaki 8528501, Japan
来源：
NUCLEIC ACIDS RESEARCH | 2020年 / 48卷 / 15期
关键词：
IN-SITU HYBRIDIZATION; MESSENGER-RNA; NUCLEOTIDE POLYMORPHISMS; CELLS; PCR; AMPLIFICATION; PROVIRUS;
D O I：
10.1093/nar/gkaa515
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
Specific nucleic acid sequences can be detected in individual cells by in situ hybridization. However, when very few copies of a target sequence are present per cell, its signal is undetectable by flow cytometry. Although various approaches have been developed to increase fluorescence signals for in situ hybridization, flow cytometric detection of specific genomic DNA sequences has not been established. Here, we present a flow cytometry assay for detection of single-copy genomic sequences in human lymphocytes using in situ PCR with universal energy transfer-labelled primers.
引用
收藏
页数：7
相关论文
共 50 条
  • [1] Continuous-Flow Polymerase Chain Reaction of Single-Copy DNA in Microfluidic Microdroplets
    Schaerli, Yolanda
    Wootton, Robert C.
    Robinson, Tom
    Stein, Viktor
    Dunsby, Christopher
    Neil, Mark A. A.
    French, Paul M. W.
    deMello, Andrew J.
    Abell, Chris
    Hollfelder, Florian
    ANALYTICAL CHEMISTRY, 2009, 81 (01) : 302 - 306
  • [2] Fluorescence Imaging of Single-Copy DNA Sequences within the Human Genome Using PNA-Directed Padlock Probe Assembly
    Yaroslavsky, Anastasia I.
    Smolina, Irina V.
    CHEMISTRY & BIOLOGY, 2013, 20 (03): : 445 - 453
  • [3] Random DNA fragmentation allows detection of single-copy, single-exon alterations of copy number by oligonucleotide array CGH in clinical FFPE samples
    Hostetter, Galen
    Kim, Su Young
    Savage, Stephanie
    Gooden, Gerald C.
    Barrett, Michael
    Zhang, Jian
    Alla, Lalitamba
    Watanabe, April
    Einspahr, Janine
    Prasad, Anil
    Nickoloff, Brian J.
    Carpten, John
    Trent, Jeffrey
    Alberts, David
    Bittner, Michael
    NUCLEIC ACIDS RESEARCH, 2010, 38 (02) : e9.1 - e9.12
  • [4] RAPID ASSESSMENT OF SINGLE-COPY NUCLEAR-DNA VARIATION IN DIVERSE SPECIES
    SLADE, RW
    MORITZ, C
    HEIDEMAN, A
    HALE, PT
    MOLECULAR ECOLOGY, 1993, 2 (06) : 359 - 373
  • [5] Generation of type-specific probes for the detection of single-copy human papillomavirus by a novel in situ hybridization method
    Huang, CC
    Qiu, JT
    Kashima, ML
    Kurman, RJ
    Wu, TC
    MODERN PATHOLOGY, 1998, 11 (10) : 971 - 977
  • [6] Single-copy nuclear DNA sequences obtained from noninvasively collected primate feces
    Surridge, AK
    Smith, AC
    Buchanan-Smith, HM
    Mundy, NI
    AMERICAN JOURNAL OF PRIMATOLOGY, 2002, 56 (03) : 185 - 190
  • [7] Flow cytometry assay for intracellular rabies virus detection
    Bordignon, J
    Ferreira, SCP
    Caporale, GMM
    Carrieri, ML
    Kotait, I
    Lima, HC
    Zanetti, CR
    JOURNAL OF VIROLOGICAL METHODS, 2002, 105 (01) : 181 - 186
  • [8] Extraction of single-copy nuclear DNA from forensic specimens with a variety of postmortem histories
    Evison, MP
    Smillie, DM
    Chamberlain, AT
    JOURNAL OF FORENSIC SCIENCES, 1997, 42 (06) : 1032 - 1038
  • [9] In situ hybridization detection of single-copy human papillomavirus on isolated cells, using a catalyzed signal amplification system:: GenPoint™
    Lizard, G
    Démares-Poulet, MJ
    Roignot, P
    Gambert, P
    DIAGNOSTIC CYTOPATHOLOGY, 2001, 24 (02) : 112 - 116
  • [10] Detection of single-copy functional genes in prokaryotic cells by two-pass TSA-FISH with polynucleotide probes
    Kawakami, Shuji
    Hasegawa, Takuya
    Imachi, Hiroyuki
    Yamaguchi, Takashi
    Harada, Hideki
    Ohashi, Akiyoshi
    Kubota, Kengo
    JOURNAL OF MICROBIOLOGICAL METHODS, 2012, 88 (02) : 218 - 223
12345