The storage period of the formalin-fixed paraffin-embedded tumor blocks does not influence the concentration and purity of the isolated DNA in a series of 83 renal and thyroid carcinomas.
نویسندگان
چکیده
Optimal recovery of nucleic acids from formalin-fixed paraffin-embedded (FFPE) tissues is highly dependent on a series of pre-extraction steps, mainly related (but not limited) to fixation. The aim of our study was to investigate if the storage period of the FFPE blocks had a significant effect on the isolated DNA. We examined the quantity and purity of the isolated DNA from 83 FFPE blocks, corresponding to malignant thyroid (n=28) and renal (n=55) carcinomas that had been stored in our department for up to eight years. The DNA extraction protocol was based on a precipitation method (MasterPure™ DNA Purification Kit, Epicentre), in accordance to the manufacturer instructions, optimized in our laboratory. A spectrophotometer was used to determine the yield (A260) and purity (A260/A280 ratio) of the isolated DNA. We successfully isolated good DNA quantity and purity from all our study cases (mean concentration: 223.4 ± 104.16 ng/μL; mean A260/A280 ratio: 1.68 ± 0.09). Moreover, no statistically significant differences were observed between tumor blocks stored for 2-3 years and 7-8 years, respectively, both in terms of DNA quantity (p=0.196) and purity (p=0.663). In conclusion, we successfully validated an efficient, reproducible DNA extraction technique that provided a good range of DNA concentrations and purity, regardless the type of tissue (thyroid or kidney). Moreover, we demonstrated that the storage period of the FFPE blocks does not have a significant influence on the DNA quantity and purity.
منابع مشابه
Molecular Identification of Mycobacterium Tuberculosis Complex in Formalin-Fixed, Paraffin-Embedded Tissue Blocks of Extra Pulmonary Speciemens using Genomics Extraction
Background: Tuberculosis has been detected in some extra pulmonary ecological niches. Although extra pulmonary tuberculosis (EPTB) is less frequent than Pulmonary Tuberculosis (PTB), its incidence has increased worldwide. The aim of this study was to investigate the presence of EPTB and MDR-EXPT in Formalin-fixed, paraffin-embedded tissue blocks among different samples in Kerma...
متن کاملExpression of Survivin in the Formalin-Fixed, Paraffin-embedded Specimens of Human Osteosarcoma May Be Used as a Diagnostic Marker
Osteosarcoma is the most frequent malignant bone tumor with a peak incidence in the second and third decades of life. Diagnosis of patients with osteosarcoma is limited to clinico-pathological parameters whereas molecular markers of tumor initiation and promotion have not yet been identified. We aimed to study the expression of the anti-apoptotic survivin gene in osteosarcoma specimens. A total...
متن کاملValue of CD10 Expression in Differentiating Cutaneous Basal from Squamous Cell Carcinomas and Basal Cell Carcinoma from Trichoepithelioma
Background: In addition to the well-defined histological criteria for squamous cell carcinoma (SCC) and basal cell carcinoma (BCC), immunohistochemical techniques can be used in difficult cases for their differentiation. As differential diagnosis between trichoepithelioma (TE) and BCC is sometimes difficult for the clinician and the pathologist, CD10 may be a useful marker for definite diagnosi...
متن کاملThe Comparison of different Procedures for DNA extraction from paraffin-embedded Tissues: A commercial kit and a traditional method based on heating
Abstract Background and objectives: Paraffin-embedded tissues and clinical samples are a valuable resource for molecular genetic studies, but the extraction of high-quality genomic DNA from this tissues is still a problematic issue. In the Present study, the efficiency of two DNA extraction protocols, a commercial kit and a traditional method based on heating and K Proteinase was compared. Mate...
متن کاملAn Efficient Method For DNA Extraction From Paraffin Wax Embedded Tissues For PCR Amplification Of Human And Viral DNA
Background and Objective: Formalin-fixed paraffin-embedded tissues are a valuable source of DNA for molecular studies. We designed and optimized an efficient procedure for DNA extraction from formalin-fixed paraffin embedded tissues. Materials and Methods: Seventy three blocks of cervical paraffin-embedded tissues were investigated. DNA was extracted using 45 minutes boiling in alkaline sol...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Romanian journal of morphology and embryology = Revue roumaine de morphologie et embryologie
دوره 56 2 Suppl شماره
صفحات -
تاریخ انتشار 2015