Representative cDNA libraries and their utility in gene expression profiling

被引:102
作者
Endege, WO
Steinmann, KE
Boardman, LA
Thibodeau, SN
Schlegel, R
机构
[1] Chiron Diagnost, E Walpole, MA USA
[2] Mayo Clin & Mayo Fdn, Rochester, MN 55905 USA
关键词
D O I
10.2144/99263cr04
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
An increasing interest in gene expression profiles in human diseases has led to the use of microdissected tumors and biopsies in gene discovery approaches. Since many of these clinical samples yield extremely small amounts of RNA, reproducible methods are needed to amplify this RNA while maintaining the original message profile. Using the SMART(TM) cDNA Synthesis Method, we show that high-, medium- and low-abundance transcripts can be amplified in a representative fashion and that the resulting cDNA can also be used as a complex probe to confirm gene expression differences identified by other techniques.
引用
收藏
页码:542 / +
页数:7
相关论文
共 15 条
  • [1] Averboukh L, 1996, PROSTATE, V29, P350
  • [2] BRADY G, 1993, METHOD ENZYMOL, V225, P611
  • [3] A RANDOM-PCR METHOD (RPCR) TO CONSTRUCT WHOLE CDNA LIBRARY FROM LOW AMOUNTS OF RNA
    FROUSSARD, P
    [J]. NUCLEIC ACIDS RESEARCH, 1992, 20 (11) : 2900 - 2900
  • [4] A SIMPLE PROCEDURE FOR GEL-ELECTROPHORESIS AND NORTHERN BLOTTING OF RNA
    GODA, SK
    MINTON, NP
    [J]. NUCLEIC ACIDS RESEARCH, 1995, 23 (16) : 3357 - 3358
  • [5] KLICKSTEIN LB, 1996, CURRENT PROTOCOLS MO, V1
  • [6] DIFFERENTIAL DISPLAY OF EUKARYOTIC MESSENGER-RNA BY MEANS OF THE POLYMERASE CHAIN-REACTION
    LIANG, P
    PARDEE, AB
    [J]. SCIENCE, 1992, 257 (5072) : 967 - 971
  • [7] Maniatis T., 1982, MOL CLONING A LAB MA
  • [8] Identification and verification of differential display cDNAs using gene-specific primers and hybridization arrays
    Martin, KJ
    Kwan, CP
    O'Hare, MJ
    Pardee, AB
    Sager, R
    [J]. BIOTECHNIQUES, 1998, 24 (06) : 1018 - +
  • [9] Elimination of false positives generated through PCR re-amplification of differential display cDNA
    Miele, G
    MacRae, L
    McBride, D
    Manson, J
    Clinton, M
    [J]. BIOTECHNIQUES, 1998, 25 (01) : 138 - 144
  • [10] Screening differentially expressed cDNA clones obtained by differential display using amplified RNA
    Poirier, GMC
    Pyati, J
    Wan, JS
    Erlander, MG
    [J]. NUCLEIC ACIDS RESEARCH, 1997, 25 (04) : 913 - 914