الصفحة الحالية:الصفحة الرئيسية
د. الشيماء الحسيني حسب النبي حسب النبي
الدرجة الوظيفية: مدرس
الوظيفة الإدارية الحالية:
الوظيفة الإدارية السابقة:
الكلية: كلية الطب البيطرى
القسم: التوليد والتناسل والتلقيح الإصطناعى
البريد الإلكتروني التعليمي: alshimaa.assabelnaby@fvtm.bu.edu.eg
البريد الإلكتروني البديل: dvet_shima@hotmail.co.uk
رقم الموبايل: 01278223318
الإسم العلمي: Al-shimaa Al-H H. El-Naby
البحوث [ عناوين(13) :: بحوث كاملة(1) :: ملخصات البحوث(13) ]
روابط (3) :: ملفات المقررات( 0) | مجموع النقاط :32
أخبار
Establishment a protocol for total RNA isolation from buffalo fresh and frozen semen for molecular applications [2020-10-20]
Abstract To date, there is no an established protocol for total RNA isolation in Egyptian buffalo spermatozoa. The present study aimed (I) to establish a defined protocol for total RNA isolation from fresh and frozen spermatozoa, (II) to evaluate RNA quality and quantity from different extraction methods and studying gene expression. Warm and standard room temperature modified QIAzol Lysis Reagents were used for total RNA extraction. The quality and quantity of extracted RNA were checked, and subsequently qRT-PCR was performed using androgen receptor-like and three reference gene primers (GAPDH, ACTB and 18S). The warm modified QIAzol Lysis Reagents resulted higher yield of good quality RNA from fresh (569.54 ± 18.83 ng/μl) and frozen spermatozoa (110.59 ± 4.43 ng/μl), compared to standard room temperature modified QIAzol (421.26 ± 7.18 ng/μl) and (29.07 ± 5.25 ng/μl), for fresh and frozen semen samples respectively. The 260/280 ratio was 1.90 and 1.89 for fresh and frozen isolated semen by warm method respectively. The integrity of RNA was good and appeared as a sharp band on 2% agarose gel. The most stable reference gene was 18S. Reliable extraction method of high quality RNA yield could be a step forward for understanding mechanisms of spermatogenesis for improving male fertility. K E Y W O R D S Egyptian buffalo, reference genes, RNA isolation, spermatozoa, warm QIAzolmoreالاهتمامات البحثية
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