معرفي يك روش جهت استخراج DNA از اكتينومايست هاي هوازي

New method for DNA extraction of the genus Nocardia

سابقه و هدف: استخراج DNA از اكتينوميست هاي هوازي به دليل پيچيدگي ساختمان ديواره سلولي آنها با روش هاي روتين براي استخراج DNA متفاوت است. در اين مطالعه از روش جوشاندن به همراه محلول STET buffer براي استخراجDNA از اين باكتري ها استفاده گرديد. مواد و روش ها: با استفاده از محلول STET buffer ، ژنوم ميكروارگانيسم ها استخراج گرديد. كيفيت و كميت DNA استخراج شده، با استفاده از اسپكتروفوتومتر و الكتروفورز بر روي ژل آگارز مورد ارزيابي قرار گرفت. يافته ها: نتايج به دست آمده از OD و الكتروفورز بر روي ژل آگارز، نشان دهنده كيفيت مناسب DNA استخراج شده جهت انجام كارهاي مولكولي بود. نتيجه گيري: ارزاني، آساني، سريع بودن و داشتن DNA ژنومي خوب از مزيت هاي اين روش مي باشد.

Aim and Background: Nocardia spp are Gram-positive, partially acid-fast causing various infections. Due to the complex structure of the cell wall of the microorganism that is similar to mycobacteria, DNA extraction from this bacterium is different from other bacteria. The boiling method with STET buffer solution was used for DNA extraction of Nocardia. Materials and Methods: Nocardia colonies were suspended in 200 µl STET buffer and was boiled for 30 minutes. Suspension was centrifuged and transferred to another micro tube and Ethanol %95 was added and saved at -20 °C for 30 minutes. After this stage, sample was centrifuged and the supernatant was discarded. In the first stage, distilled water was added and stored at -20 °C for molecular works. To confirm the presence and quality of the DNA extract, the electrophoresis on a %1 agarose gel was carried out and PCR reaction 16S rRNA gene was used. Results: DNA was extracted on agarose gel and the purity and quality was appropriate. 16S rRNA gene PCR was then performed and observed to be 1500 bp