FORMALIN-INDUCED INFIDELITY IN PCR-AMPLIFIED DNA FRAGMENTS

被引:32
作者
DEGIORGI, C [1 ]
SIALER, MF [1 ]
LAMBERTI, F [1 ]
机构
[1] CNR,IST NEMATOL AGRARIA,I-70126 BARI,ITALY
关键词
ARTIFACTS; FORMALIN; NEMATODE; PCR; SEQUENCE;
D O I
10.1006/mcpr.1994.1065
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A 643-nucleotide-long fragment of rDNA gene was amplified by PCR in the nematode worm Caenorhabditis elegans. When the experiments were performed by using samples fixed in formalin, artefacts were detected. While the size of the amplified fragment resulted unaffected, very striking differences were seen in the nucleotide sequences of the amplified fragments. Furthermore, in many cases, the PCR reaction failed completely. The results obtained might warn of potential problems, especially when the amount of DNA to be amplified is scarce.
引用
收藏
页码:459 / 462
页数:4
相关论文
共 14 条
  • [1] HISTONE-HISTONE PROPINQUITY BY ALDEHYDE FIXATION OF CHROMATIN
    CHALKLEY, R
    HUNTER, C
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1975, 72 (04) : 1304 - 1308
  • [2] THE RDNA OF C-ELEGANS - SEQUENCE AND STRUCTURE
    ELLIS, RE
    SULSTON, JE
    COULSON, AR
    [J]. NUCLEIC ACIDS RESEARCH, 1986, 14 (05) : 2345 - 2364
  • [3] DEPURINATION-INDUCED INFIDELITY OF DEOXYRIBONUCLEIC-ACID SYNTHESIS WITH PURIFIED DEOXYRIBONUCLEIC-ACID REPLICATION PROTEINS INVITRO
    KUNKEL, TA
    SCHAAPER, RM
    LOEB, LA
    [J]. BIOCHEMISTRY, 1983, 22 (10) : 2378 - 2384
  • [4] INSTABILITY AND DECAY OF THE PRIMARY STRUCTURE OF DNA
    LINDAHL, T
    [J]. NATURE, 1993, 362 (6422) : 709 - 715
  • [5] NUOVO GJ, 1991, AM J PATHOL, V139, P847
  • [6] POLYMERASE CHAIN-REACTION REVEALS CLONING ARTIFACTS
    PAABO, S
    WILSON, AC
    [J]. NATURE, 1988, 334 (6181) : 387 - 388
  • [7] ANCIENT DNA - EXTRACTION, CHARACTERIZATION, MOLECULAR-CLONING, AND ENZYMATIC AMPLIFICATION
    PAABO, S
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (06) : 1939 - 1943
  • [8] PAABO S, 1990, J BIOL CHEM, V265, P4718
  • [9] PRIMER-DIRECTED ENZYMATIC AMPLIFICATION OF DNA WITH A THERMOSTABLE DNA-POLYMERASE
    SAIKI, RK
    GELFAND, DH
    STOFFEL, S
    SCHARF, SJ
    HIGUCHI, R
    HORN, GT
    MULLIS, KB
    ERLICH, HA
    [J]. SCIENCE, 1988, 239 (4839) : 487 - 491
  • [10] INFIDELITY OF DNA-SYNTHESIS ASSOCIATED WITH BYPASS OF APURINIC SITES
    SCHAAPER, RM
    KUNKEL, TA
    LOEB, LA
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1983, 80 (02): : 487 - 491