Kullmann, F. and Bocker, T. and Schölmerich, Jürgen and Rüschoff, J. (1998) A comparison of methods for DNA extraction from paraffin-embedded tissue for microsatellite instability analysis by polymerase chain reaction. ACTA BIOTECHNOLOGICA, 18 (1). pp. 77-83. ISSN 0138-4988
Full text not available from this repository.Abstract
In a prospective study, nuclear DNA was extracted from colorectal tumours and normal mucosa which had been fixed in buffered formalin and embedded into paraffin. DNA-extraction was performed using three different methods: a commercial kit which was not especially created for this use; a known fast procedure without DNA-cleaning steps; and a more conventional DNA-preparation protocol with DNA-cleaning. Using the polymerase chain reaction (PCR), DNA was amplified by being targeted onto two beta-globin fragments with different lengths (536 bp and 989 bp) and (CA)n repeats localized on chromosome 5q (D5S346) and chromosome 17p (TP53CA) with a length of about 100bp for detection of microsatellite instability. The success rate of microsatellite amplification was 100% with all methods. The 536 bp beta-globin fragment could be amplified with a success rate ranging from 40% to 100%. The amplification of the 989 bp beta-globin fragment was unsuccessful. Significant differences were observed between the three methods in the final DNA concentration and DNA yield. In microsatellite instability studies of paraffin-embedded tissues, the investigator can expect a high success rate of nearly 100% using any of the described methods.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | AMPLIFICATION |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Innere Medizin I Medicine > Lehrstuhl für Pathologie |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 28 Mar 2023 04:42 |
| Last Modified: | 28 Mar 2023 04:42 |
| URI: | https://pred.uni-regensburg.de/id/eprint/50160 |
Actions (login required)
 |
View Item |
