Método para proveer fragmentos de ADN derivados de una muestra archivada

• Main Authors: WASSERKORT, Reinhold, DIETRICH, Dimo, SCHUSTER, Matthias, KLUTH, Antje, BALLHAUSE, Matthias, WAGNER, Ute, ZIEBARTH, Heike, BERLIN, Kurt
• Format: Patent
• Language: Spanish
• Published: 16.10.2020
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; COMPOSITIONS THEREOF; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; CULTURE MEDIA; ENZYMOLOGY; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

Un método para amplificar el ADN tratado con bisulfito derivado de una muestra archivada por reacción en cadena de polimerasa, que comprende amplificar el ADN tratado con bisulfito en una etapa de amplificación de ADN que comprende el uso de una concentración de polimerasa en el rango de 0.05 a 0.3 U/μl y una concentración de cada nucleótido en el rango de 350 a 650 μmol/l. Aspects of the present invention relate to compositions and methods for providing DNA fragments from an archived sample (e.g., paraffin-embedded and/or fixed-tissue biopsies, etc). Particular aspects provide methods whereby high yields of DNA are isolated as well as a substantial portion of the DNA consists of long DNA fragments, and where the isolated genomic DNA is free of associated or cross-linked contaminants like proteins, peptides, amino acids or RNA. The methods are facile, cost-effective, an are characterized by high reproducibility and reliability. Particular aspects provide methods for providing DNA fragments derived from an archived sample, wherein the yield of DNA before, for example, an amplification step is at least 20%, and amplicons up to a length of about 1,000 base pairs are amplifiable.