Evaluation of the automated fluorescent analysis of the H-ras minisatellite after optimized PCR amplification in comparison with a standardized Southern blot technique

  • Authors:
    • S Gosse
    • S Sauvaigo
    • A Daver
    • F Larra
    • J Saavedra
    • J Marchand
    • D BernardGallon
    • Y Bignon
  • View Affiliations

  • Published online on: April 1, 1997     https://doi.org/10.3892/ijo.10.4.735
  • Pages: 735-740
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Abstract

Determination of allele sizes? loss of heterozygosity or genetic instability at minisatellite VNTR loci, are routinely performed by the conventional Southern technique. We have investigated the potential use of automated DNA sequencer for the analysis of the H-ras minisatellite. We report the modifications of amplification parameters and electrophoresis conditions on the sequencer. Seventy-one colorectal carcinomas and the corresponding normal tissues were amplified with fluorescent-labeled primers, analyzed on sequencer, and concurrently controlled by Southern blotting. The results on sequencer showed that a Hydrolink matrix used in non-denaturing conditions and a specific analysis software facilitate a more accurate fragment size calculation.

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April 1997
Volume 10 Issue 4

Print ISSN: 1019-6439
Online ISSN:1791-2423

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Spandidos Publications style
Gosse S, Sauvaigo S, Daver A, Larra F, Saavedra J, Marchand J, BernardGallon D and Bignon Y: Evaluation of the automated fluorescent analysis of the H-ras minisatellite after optimized PCR amplification in comparison with a standardized Southern blot technique. Int J Oncol 10: 735-740, 1997
APA
Gosse, S., Sauvaigo, S., Daver, A., Larra, F., Saavedra, J., Marchand, J. ... Bignon, Y. (1997). Evaluation of the automated fluorescent analysis of the H-ras minisatellite after optimized PCR amplification in comparison with a standardized Southern blot technique. International Journal of Oncology, 10, 735-740. https://doi.org/10.3892/ijo.10.4.735
MLA
Gosse, S., Sauvaigo, S., Daver, A., Larra, F., Saavedra, J., Marchand, J., BernardGallon, D., Bignon, Y."Evaluation of the automated fluorescent analysis of the H-ras minisatellite after optimized PCR amplification in comparison with a standardized Southern blot technique". International Journal of Oncology 10.4 (1997): 735-740.
Chicago
Gosse, S., Sauvaigo, S., Daver, A., Larra, F., Saavedra, J., Marchand, J., BernardGallon, D., Bignon, Y."Evaluation of the automated fluorescent analysis of the H-ras minisatellite after optimized PCR amplification in comparison with a standardized Southern blot technique". International Journal of Oncology 10, no. 4 (1997): 735-740. https://doi.org/10.3892/ijo.10.4.735