An Efficient Method for DNA Extraction from Paraffin Wax Embedded Tissues for PCR Amplification of Human and Viral DNA
Publish place: IRANIAN JOURNAL of PATHOLOGY، Vol: 3، Issue: 4
Publish Year: 1387
نوع سند: مقاله ژورنالی
زبان: English
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شناسه ملی سند علمی:
JR_IJP-3-4_011
تاریخ نمایه سازی: 5 آبان 1393
Abstract:
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 extractionfrom formalin-fixed paraffin embedded tissues. Materials and Methods: Seventy three blocks of cervical paraffin-embedded tissues wereinvestigated. DNA was extracted using 45 minutes boiling in alkaline solution together with 10 beads of Chelex-20, followed by phenol-chloroform extraction and alcohol precipitation. Results: This method produced DNA suitable for amplification using primers specific for humanSMN and β globin genes in 98.63% and 82.2% of samples respectively. We also detected human papillomavirus DNA in 58.33% of appropriate samples.Conclusion: This procedure provides simple and efficient method for recovery of amplifiable genomic and viral DNA from paraffin wax embedded tissues.
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Authors
Bahare Tabanifar
Dept. of Microbiology, School of Biology, Alzahra University, Tehran, Iran
Rasoul Salehi
Dept. of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences,Isfahan, Iran
Ezat Asgarani
Dept. of Genetics, School of Biology, Alzahra University, Tehran, Iran
Mehri Faghihi
Dept. of Pathology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran