PCR AMPLIFICATION OF 40-YEAR-OLD PARAFFIN-EMBEDDED TUMOR-TISSUES - COMPARISON OF 4 DIFFERENT DNA EXTRACTION AND PURIFICATION METHODS

  • Authors:
    • WG WANG
    • P KUMAR
    • M SCHWARZ
    • G MALONE
    • A HAWORTH
    • S KUMAR
  • View Affiliations

  • Published online on: September 1, 1994     https://doi.org/10.3892/ijo.5.3.453
  • Pages: 453-457
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Abstract

Four different methods of DNA extraction from formalin-fixed, paraffin-embedded tissues were compared for their ability to produce DNA suitable as a template for polymerase chain reaction (PCR). Seven paraffin-embedded rhabdomyosarcoma tissue blocks from the 1950s and one from 1960 were treated with the following extraction methods: 1. Proteinase K digestion and phenol/chloroform extraction; 2. Proteinase K digestion followed by boiling to inactivate the enzyme; 3. Proteinase K digestion, addition of Chelex-100, followed by boiling; and 4. Proteinase K digestion and Prep-A-Gene purification. DNA extracted by methods 1 and 2 was degraded, but DNA of high molecular weight was recovered in every sample extracted by methods 3 and 4 - even though some degradation was observed. Extracted DNA was used as a template for PCR amplification of exon 4 of the PAX-3 gene. PCR was successful in 7 out of 8 samples prepared using methods 3 and 4, producing levels of amplified product equivalent to those obtained with control DNA obtained from fresh lymphocytes. Only very weak products were found in samples prepared by methods 1 (2 out of 8) and 2 (4 out of 8). These results indicate that chelation of polyvalent ions (Chelex-100 method) or obviating the need for boiling (Prep-A-Gene) may protect DNA during extraction.

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September 1994
Volume 5 Issue 3

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

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Spandidos Publications style
WANG W, KUMAR P, SCHWARZ M, MALONE G, HAWORTH A and KUMAR S: PCR AMPLIFICATION OF 40-YEAR-OLD PARAFFIN-EMBEDDED TUMOR-TISSUES - COMPARISON OF 4 DIFFERENT DNA EXTRACTION AND PURIFICATION METHODS. Int J Oncol 5: 453-457, 1994
APA
WANG, W., KUMAR, P., SCHWARZ, M., MALONE, G., HAWORTH, A., & KUMAR, S. (1994). PCR AMPLIFICATION OF 40-YEAR-OLD PARAFFIN-EMBEDDED TUMOR-TISSUES - COMPARISON OF 4 DIFFERENT DNA EXTRACTION AND PURIFICATION METHODS. International Journal of Oncology, 5, 453-457. https://doi.org/10.3892/ijo.5.3.453
MLA
WANG, W., KUMAR, P., SCHWARZ, M., MALONE, G., HAWORTH, A., KUMAR, S."PCR AMPLIFICATION OF 40-YEAR-OLD PARAFFIN-EMBEDDED TUMOR-TISSUES - COMPARISON OF 4 DIFFERENT DNA EXTRACTION AND PURIFICATION METHODS". International Journal of Oncology 5.3 (1994): 453-457.
Chicago
WANG, W., KUMAR, P., SCHWARZ, M., MALONE, G., HAWORTH, A., KUMAR, S."PCR AMPLIFICATION OF 40-YEAR-OLD PARAFFIN-EMBEDDED TUMOR-TISSUES - COMPARISON OF 4 DIFFERENT DNA EXTRACTION AND PURIFICATION METHODS". International Journal of Oncology 5, no. 3 (1994): 453-457. https://doi.org/10.3892/ijo.5.3.453