Abstract This study is to identify some mutations in exons 23, 24, 26 A+B, and 26 A of the factor VIII gene that causes hemophilia A among Libyan patients. Forty-three patients (40 males, 3 females), 10 carrier, and 10 control (8 males, 2 females) were enrolled in this study. The total number were 63 samples for each exon. Blood samples were collected from Libyan patients (Department of Paediatric Hematology, Tripoli Medical Center), and kept in freezer at -20 Cº. The blood samples were transported to DNA laboratory in the Genetic Engineering Department and Human Tissues Department Biotechnology Research Center, Twesha- Libya. Genomic DNA was extracted from whole blood and DNA was visualized by Agarose Gel Electrophoresis, and DNA concentration was measured by Spectrophotometer. DNA was amplified by polymerase chain reaction (PCR) technique. Mutations were identified by restriction enzymes Taq I and Xba I. Taq I was used to detect nonsense mutations in Exon 23, 2147 [ Arg Stop codon (Term)], Exon 24, 2209 [ Arg Stop codon (Term)], and Exon 26 A+B, 2307 [ Arg Stop codon (Term)]. Our result showed no mutations were detected by using restriction enzyme Taq I in Exons 23, codon 2147, Exon 24, codon 2209, and Exon 26A+B, codon 2307. But by using restriction enzyme Xba I, we found six insertion mutations in Exon 26A, 3919 (ins GA) 11.3 % of 53 patients, and two partial deletion mutations in Exon 23 (3.8 % of 53 patients) refer to area failed to be amplified by PCR.
نادية نور الدين الصربوط (2010)

Efficient extraction of genomic DNA (gDNA) from biological materials found in harsh environments is the first step for successful forensic DNA profiling. This study aimed to evaluate two methods for DNA recovery from animal tissues (livers, muscles), focusing on the best storage temperature for DNA yield in term of quality, quantity, and integrity for use in several downstream molecular techniques. Six male Swiss albino mice were sacrificed, liver and muscle tissues (n=32) were then harvested and stored for one week in different temperatures, -20C, 4C, 25C and 40C. The conditioned animal tissues were used for DNA extraction by Chelex-100 method or NucleoSpin Blood and Tissue kit. The extracted gDNA was visualized on 1.5% agarose gel electrophoresis to determine the quality of gDNA and analysed spectrophotometrically to determine the DNA concentration and the purity. Both methods, Chelex-100 and NucleoSpin Blood and Tissue kit found to be appropriate for yielding high quantity of gDNA, with the Chelex100 method yielding a greater quantity (P < 0.045) than the kit. At -20C, 4C, and 25C temperatures, the concentration of DNA yield was numerically lower than at 40C. The NucleoSpin Blood and Tissue kit produced a higher (P=0.031) purity product than the Chelex-100 method, particularly for muscle tissues. The Chelex-100 method is cheap, fast, effective, and is a crucial tool for yielding DNA from animal tissues (livers, muscles) exposed to harsh environment with little limitations.
Huda H. Al-Griw, Zena A. Zraba, Salsabiel K. Al-Muntaser, Marwan M. Draid, Aisha M. Zaidi, Refaat M. Tabagh , Mohamed A. Al-Griw(8-2017)

سرطان الثدي من أكثر الأمراض خبثا، ويعتبر المسبب الأساسي للوفاة بين النساء بعد سرطان الرئة. وهو من أكثر الأمراض انتشارا بين النساء بعد سرطان الجلد. والهدف من هذه الدراسة هو التعرف على احتمال وجود الطفرتين 185delAG و5382insCلمورثة سرطان الثدي1 (BRCA1) بين 77 سيدة ليبية من اللواتي يترددن على عيادة الثدي-بقسم الجراحة – المستشفى المركزي-طرابلس – ليبيا. والحالات المختارة كانت: 32 حالة مشخصة بسرطان الثدي و8 حالات من المصابات بأمراض حميدة بالثدي , 24 حالة من أقارب المريضات بسرطان الثدي، و13 سيدة غير مصابة بسرطان الثدي كمجموعة السيطرة. تم سحب خمسة مليليتر من الدم من كل امرأة ، وجمعت في أنابيب تحتوي على مادة مانعة للتجلط الدم (EDTA) وحفظت في المجمد في (-20ºC) ، نقلت عينات الدم إلى معامل الأحماض النووية لقسم الأنسجة البشرية وقسم الهندسة الوراثية – مركز بحوث التقنيات الحيوية – طويشة – ليبيا ، لأجل دراسة الحمض النووي ، باستخدام سلسلة تفاعل إنزيم متعدد البلمرة (Polymerase Chain Reaction (PCR)) لكشف الطفرات بواسطة سلسلة تفاعل إنزيم متعدد البلمرة مفصولة الطفرات )((MS-PCR Mutagenically Separated PCR) لتضخيم exons الطبيعية و exons المطفرة وذلك بواسطة بوادئ ((Primers خاصة لتشخيص الطفرات , إذا عُثر على الطفرة في أحد exons سيكون هناك حزمتان ((band باستخدام الترحيل الكهربي الهلامي (Agarose gel Electrophoresis) لاكتشاف الطفرات في exons2 و20. نحن وُصِلنَا إلى الهدفِ في هذه الدراسةِ، لمعْرِفة الوجودِ أَو عدمِ الوجود لطفراتْ delAG185 و5382insC في مورثة BRCA1 بين الإناث الليبياتِ، اللواتي يترددن على عيادةَ الثدي-قسم الجراحةِ-مستشفى المركزي-طرابلس-ليبيا؛ كَانَ كالتّالي: -الطفرة 185delAG كَانَت موجودة في 52 من أصل 77 (67.5 %) مِنْ العينات المدروسة، والطفرة 5382insC كَانَت غائبَة في هذه الدراسةِ، حيث العينات لَها حزمة واحدة (تكون حزمة طبيعيةَ) بينما العيناتَ لَها حزمتان (تكون حزمة واحدة طبيعيةَ والحزمة الأخرى غير طبيعية). وفي الخلاصة، نَتائِجنا كانت مشابهة لنَتائِجِ الدِراساتِ الأخرى، حيث تُساعدُ معرفةَ وجودِ الطفرات في التشخيصِ المبكّرِ وهذه تَزِيدُ معدل بقاء على قيد الحياة للنِساءِ للمصابات بسرطان الثدي. Abstract Breast cancer is more the malignancies diseases, and is the primary cause of death among women after lung cancer; is the most spread diseases among women with the exception of skin cancer. The aim of this study is to identify the presence of mutations 185delAG and 5382insC for BRCA1gene in 77 Libyan females attending the breast clinic, Surgery Department, Central Hospital, Tripoli - Libya. The selected cases were: 32 cases diagnosed with breast cancer and 8 cases of women with benign breast disease, 24 cases of relatives of patients with breast cancer, and 13 women without breast cancer as control. Five milliliters of blood were drawn from each woman, and collected in tubes containing EDTA anticoagulant and kept in freezer (-20ºC). The blood samples were transported to DNA laboratory for the Human's Tissues Department and the Genetic Engineering Department- Biotechnology Research Centre-Twesha - Libya, for DNA study; using Polymerase Chain Reaction (PCR); to detect mutations by Mutagenically Separated PCR (MS -PCR), to amplify the normal exons and mutant exons by specific primers for diagnosis of these mutations; if the mutation was found in one of the exons; there will be two bands by use Agarose gel Electrophoresis to detect mutations in exons 2 and 20.We have been reached to the aim this study, for knowing the existence or non-existence of two mutations,185delAG and 5382insC in BRCA1 gene among Libyan females attending the breast clinic, Surgery Department, Central Hospital, Tripoli-Libya; were as follows:- the 185delAG mutation was present in 52 out of 77 (67.5%) of studied samples, and 5382insC mutation was absent in this study, where samples have one band (are normal band) while samples have two bands ( are normal band & another band abnormal). In conclusion, our results were similar to the results of other studies, where knowledge of the existence of mutations helps in early diagnosis and this increases the survival rate for women with breast cancer.
أمل عبد السلام الصربوط (2011)