Highly sensitive method to detect mRNAs in individual cells by direct RT-PCR using Tth DNA polymerase

被引:14
作者
Chiocchia, G [1 ]
Smith, KA [1 ]
机构
[1] NEW YORK HOSP,CORNELL MED CTR,NEW YORK,NY 10021
关键词
D O I
10.2144/97222st04
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A new method for detecting the expression of low-abundance mRNA molecules has been developed that combines the sensitivity of PCR, the high efficiency and specificity of reverse transcription (RT) using Tth DNA polymerase at high temperature, and the enhancement of sensitivity and specificity of nested PCR. This method is highly sensitive, reproducible and allows the detection of MRNAs in individual cells by direct RT-PCR.
引用
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页码:312 / &
页数:4
相关论文
共 8 条
  • [1] FUNCTIONAL ISOLATION AND CHARACTERIZATION OF HUMAN HEMATOPOIETIC STEM-CELLS
    BERARDI, AC
    WANG, AL
    LEVINE, JD
    LOPEZ, P
    SCADDEN, DT
    [J]. SCIENCE, 1995, 267 (5194) : 104 - 108
  • [2] KUMAZAKI T, 1994, BIOTECHNIQUES, V16, P1017
  • [3] CO-ENGAGEMENT OF CD3 WITH LFA-1 OR ICAM-1 ADHESION MOLECULES ENHANCES THE FREQUENCY OF ACTIVATION OF SINGLE MURINE CD4+ AND CD8+ T-CELLS AND INDUCES SYNTHESIS OF IL-3 AND IFN-GAMMA BUT NOT IL-4 OR IL-6
    MARASKOVSKY, E
    TROUTT, AB
    KELSO, A
    [J]. INTERNATIONAL IMMUNOLOGY, 1992, 4 (04) : 475 - 485
  • [4] OBRIEN DP, 1994, BIOTECHNIQUES, V16, P586
  • [5] NOVEL METHOD FOR STUDYING MESSENGER-RNA PHENOTYPES IN SINGLE OR SMALL NUMBERS OF CELLS
    RAPPOLEE, DA
    WANG, A
    MARK, D
    WERB, Z
    [J]. JOURNAL OF CELLULAR BIOCHEMISTRY, 1989, 39 (01) : 1 - 11
  • [6] SCHRIEVER F, 1991, BLOOD, V77, P787
  • [7] Uberla K, 1991, PCR Methods Appl, V1, P136
  • [8] ZIEGLER BL, 1992, BIOTECHNIQUES, V13, P726