The addition of low numbers of 3′ thymine bases can be used to improve the hybridization signal of oligonucleotides for use within arrays on nylon supports

被引:33
作者
Brown, TJ [1 ]
Anthony, RM
机构
[1] Guys & St Thomas Hosp Trust, St Thomas Hosp, Dept Microbiol, London SE1 7EH, England
[2] Univ London Kings Coll, St Thomas Hosp, Dept Microbiol, London SE1 7EH, England
关键词
array; hybridisation; nylon membrane; oligonucleotide; poly dT;
D O I
10.1016/S0167-7012(00)00175-5
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Oligonucleotide arrays can be used for the analysis of microbial nucleic acid. The addition of high numbers of dTTP to the 3' ends of oligonucleotides using terminal transferase has been shown to facilitate membrane binding. This paper demonstrates low numbers of thymine bases added to the 3' end of oligonucleotides during synthesis can improve hybridisation signal intensity where the signal seen with the unmodified oligonucleotides is poor. Thus, the addition of variable numbers of thymine bases to different oligonucleotides allows the production of oligonucleotide arrays producing strong interpretable hybridisation signals. (C) 2000 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:203 / 207
页数:5
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共 8 条
  • [1] Rapid diagnosis of bacteremia by universal amplification of 23S ribosomal DNA followed by hybridization to an oligonucleotide array
    Anthony, RM
    Brown, TJ
    French, GL
    [J]. JOURNAL OF CLINICAL MICROBIOLOGY, 2000, 38 (02) : 781 - 788
  • [2] RAPID DETECTION OF RIFAMPICIN RESISTANCE IN SPUTUM AND BIOPSY SPECIMENS FROM TUBERCULOSIS PATIENTS BY PCR AND LINE PROBE ASSAY
    DEBEENHOUWER, H
    LHIANG, Z
    JANNES, G
    MIJS, W
    MACHTELINCKX, L
    ROSSAU, R
    TRAORE, H
    PORTAELS, F
    [J]. TUBERCLE AND LUNG DISEASE, 1995, 76 (05): : 425 - 430
  • [3] DNA chips: promising toys have become powerful tools
    Gerhold, D
    Rushmore, T
    Caskey, CT
    [J]. TRENDS IN BIOCHEMICAL SCIENCES, 1999, 24 (05) : 168 - 173
  • [4] GENETIC-ANALYSIS OF AMPLIFIED DNA WITH IMMOBILIZED SEQUENCE-SPECIFIC OLIGONUCLEOTIDE PROBES
    SAIKI, RK
    WALSH, PS
    LEVENSON, CH
    ERLICH, HA
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (16) : 6230 - 6234
  • [5] Steric factors influencing hybridisation of nucleic acids to oligonucleotide arrays
    Shchepinov, MS
    CaseGreen, SC
    Southern, EM
    [J]. NUCLEIC ACIDS RESEARCH, 1997, 25 (06) : 1155 - 1161
  • [6] GENUS-SPECIFIC POLYMERASE CHAIN-REACTION FOR THE MYCOBACTERIAL DNAJ GENE AND SPECIES-SPECIFIC OLIGONUCLEOTIDE PROBES
    TAKEWAKI, S
    OKUZUMI, K
    ISHIKO, H
    NAKAHARA, K
    OHKUBO, A
    NAGAI, R
    [J]. JOURNAL OF CLINICAL MICROBIOLOGY, 1993, 31 (02) : 446 - 450
  • [7] Genetic organization of the mecA region in methicillin-susceptible and methicillin-resistant strains of Staphylococcus sciuri
    Wu, SW
    de Lencastre, H
    Tomasz, A
    [J]. JOURNAL OF BACTERIOLOGY, 1998, 180 (02) : 236 - 242
  • [8] The importance of coulombic end effects:: Experimental characterization of the effects of oligonucleotide flanking charges on the strength and salt dependence of oligocation (L8+) binding to single-stranded DNA oligomers
    Zhang, WT
    Ni, HH
    Capp, MW
    Anderson, CF
    Lohman, TM
    Record, MT
    [J]. BIOPHYSICAL JOURNAL, 1999, 76 (02) : 1008 - 1017