A new DNA extraction protocol for formalin-fixed paraffin embedded tissue of uterus cancer
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Abstract (EN)
Formalin-fixed paraffin-embedded (FFPE) tissue is one of the best ways to store a material for further molecular analysis. Further molecular analyses are needed to confirm the diagnosis in such tumors like endometrial tumors. That's why, molecular analyses are important in the anatomical pathology. Extracting DNA from FFPE material is the first step leading to molecular analyses. On the other it is still challenging. Different protocols have been developed to solve this problem. Even some commercial isolation kits are available which are specialized for FFPE tissues. All of the methods aiming to develop efficient and high quality DNA for further analyses. In this study, we aimed to develop a new DNA extraction method from 30 FFPE of uterus tumor tissue blocks which were obtained with permission of Harran University Research Hospital, Department of Pathology. The quantity and quality of the extracted DNA from three replicates of each block were measured by spectrophotometer. The average quality of DNA was found between 1.7-2.0 (A260/A280) and the average absorbance ratio of A260/A230 was 1.39. The average DNA yield was 40.4 nano gram per microliter. The quality and quantity results of extracted DNA from FFPE tissues were enough for molecular analysis. The new DNA extraction protocol has many advantages such as small amount of FFPE tissues (30 micro grams per sample), six easy steps with common chemicals and solutions (without expensive chemicals such as Proteinase K and RNase A), time efficiency (extraction of 30 samples less than three hours) and cost-effective (less than 1 US dollar per sample). The new DNA extraction protocol can be adapted to every laboratory which works on DNA based technologies.
Author
Ragaz Azad Hamad Hamad
How to Cite
Ragaz Azad Hamad Hamad (Master Thesis). A new DNA extraction protocol for formalin-fixed paraffin embedded tissue of uterus cancer, 2024, Harran University.
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