Zen Hafy, V. Larasati, Riana Puspita, S Novizar, M. Haekal, A. Rafdi, R Sentani
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引用次数: 0
摘要
福尔马林固定石蜡包埋(FFPE)档案组织是目前分子研究的一种现成的资源。FFPE组织中提取的核酸,特别是线粒体DNA (mtDNA)的质量受贮存时间的影响。因此,本研究调查了FFPE组织的储存时间是否对提取mtDNA的质量有影响,在RSUP病理学部Mohammad Hoesin Palembang博士。DNA是从RSUP Mohammad Hoesin Palembang博士病理解剖实验室随机选择的16个档案FFPE组织中提取的。根据贮藏时间(1年以下和1 ~ 5年)进行分组。“老)。用PCR方法扩增各组分离的DNA,分别设计两对引物,扩增长度分别为320 bp和142bp的mtDNA。用电泳法观察PCR产物。320bp引物均不能扩增两组样本。然而,每个研究组的149bp引物的PCR结果均为阳性。该研究表明,储存时间不会影响保存在RSUP Mohammad Hoesin Palembang博士病理学部的FFPE样本中分离的mtDNA的质量。此外,研究表明,从FFPE组织中提取的mtDNA已经降解,因此,样品不适合用于需要长mtDNA扩增子的遗传学研究。
Hubungan Lama Penyimpanan Sampel Arsip Jaringan Dalam Blok Parafin Terfiksasi Formalindengan Kualitas Hasil Ekstraksidna Mitokondria Jaringan
Formalin-fixed paraffin-embedded (FFPE) archival tissue presents a readily available resource in molecular study nowadays. The quality of nucleid acid, especially mitoconhdrial DNA (mtDNA) extracted from FFPE tissue could be affected by the storage time. Thus, this study investigated if the FFPE tissue’s storage time had an effect on the quality of the extracted mtDNA at Department of Pathology RSUP Dr. Mohammad Hoesin Palembang. DNA was extracted from 16 randomly selected archival FFPE tissues in Laboratory of Pathology Anatomy, RSUP dr. Mohammad Hoesin Palembang. The samples were grouped based on their storage time (less than 1 year and 1 to 5 yrs.’ old). The isolated DNA from each group was amplified using PCR with two primer pairs specifically designto amplify mtDNA of 320 bp and 142bp length, respectively. The PCR products were visualized by electrophoresismethod. None of the samples from both groups could be amplified witht the 320bp primers. However, the PCR result of the 149 bp primers showed positive for all of the samples from each study group. The study indicated that the storage time does not affect the quality of mtDNA isolated from the FFPE samples archived in Department of Pathology RSUP Dr. Mohammad Hoesin Palembang. Furthermore, the study showed that the mtDNA extracted from FFPE tissue has been degraded, therefore, the samples are not suitable for genetic studies require a long mtDNA amplicon.