Comparison of Methods for the Extraction of DNA from Formalin-Fixed, Paraffin-Embedded Archival Tissues

نویسندگان

  • Burcu Sengüven
  • Emre Baris
  • Tulin Oygur
  • Mehmet Berktas
چکیده

AIM Discussing a protocol involving xylene-ethanol deparaffinization on slides followed by a kit-based extraction that allows for the extraction of high quality DNA from FFPE tissues. METHODS DNA was extracted from the FFPE tissues of 16 randomly selected blocks. Methods involving deparaffinization on slides or tubes, enzyme digestion overnight or for 72 hours and isolation using phenol chloroform method or a silica-based commercial kit were compared in terms of yields, concentrations and the amplifiability. RESULTS The highest yield of DNA was produced from the samples that were deparaffinized on slides, digested for 72 hours and isolated with a commercial kit. Samples isolated with the phenol-chloroform method produced DNA of lower purity than the samples that were purified with kit. The samples isolated with the commercial kit resulted in better PCR amplification. CONCLUSION Silica-based commercial kits and deparaffinized on slides should be considered for DNA extraction from FFPE.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

An 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 کامل

The 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 کامل

Recovery of ranavirus dsDNA from formalin-fixed archival material.

The extraction and amplification of nucleic acid from formalin-fixed and paraffin-embedded tissues has become an important exercise in the collection of retrospective epidemiological data. A protocol is described that enables the extraction and amplification of dsDNA from fixed tissues within paraffin blocks and from specimens stored in 10% (aq) formalin. The procedure can be used for the exami...

متن کامل

Comparison of methods in the recovery of nucleic acids from archival formalin-fixed paraffin-embedded autopsy tissues.

Archival formalin-fixed paraffin-embedded (FFPE) human tissue collections are typically in poor states of storage across the developing world. With advances in biomolecular techniques, these extraordinary and virtually untapped resources have become an essential part of retrospective epidemiological studies. To successfully use such tissues in genomic studies, scientists require high nucleic ac...

متن کامل

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 کامل

Molecular detection and genotypic characterization of Toxoplasma gondii in paraffin-embedded fetoplacental tissues of women with recurrent spontaneous abortion

Background Congenital toxoplasmosis is an important cause of spontaneous abortion worldwide. However, there is limited information on detection and genotypic characterization of T. gondii in women with recurrent spontaneous abortion (RSA). The aim of this study is molecular detection and genotypic characterization of T. gondii in formalin-fixed, paraffin-embedded fetoplacental tissues (FFPTs) o...

متن کامل

ذخیره 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 نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:

دوره 11  شماره 

صفحات  -

تاریخ انتشار 2014