| Autor: |
Chia BH, Ernie Suhaiza BR, Syarifah Aqlimi SMZ, Norhafizah BCAR, Sasongko TH, Che Anuar CY |
| Zdroj: |
International Medical Journal; Mar2012, Vol. 19 Issue 1, p7-10, 4p, 1 Diagram, 2 Charts |
| Abstrakt: |
Objective: Pure and high throughput DNA isolation from amniotic fluid is a major bottleneck for most molecular genetic studies as well as diagnostic services. The objective of this study was to develop an in-house protocol for isolating DNA from amniotic fluid using salting-out procedure. Study design: Sixty amniotic fluid specimens from healthy pregnant women were collected after an informed consent. DNA were extracted using salting-out procedure followed by ethanol precipitation. Successful isolation was determined by purity, concentration and usability assessment. Purity and concentration of the extracted DNA was measured using nano-spectrophotometer. Usability assessment was performed using PCR for amplifying CFTR and SRY genes. Second cycle of ethanol precipitation was done for DNA samples which did not pass usability assessment. Results: DNA was successfully isolated from all amniotic fluid specimens after at least two cycles of ethanol precipitations. Conclusion: We have developed an in-house protocol for extracting DNA from amniotic fluid. We proposed a workflow protocol for such purpose and suggested that two-cycle ethanol precipitation may be necessary for DNA extracted from amniotic fluid using salting-out procedure. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Supplemental Index |
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