Document Details

Document Type : Thesis 
Document Title :
QUALITY ASSESSMENT AND UTILIZATION OF RNA EXTRACTED FROM IMMUNOSTAINED FORMALIN FIXED PARAFFIN EMBEDDED (FFPE) SLIDES IN MOLECULAR CANCER BIOLOGY
تقييم جودة الحمض النووي الريبي المستخرج من عينات الشرائح الشمعية المصبوغة كيمائيا ومناعيا وتطبيقاته لدراسة البيولوجيا الجزيئية للسرطان
 
Subject : Faculty of Science 
Document Language : Arabic 
Abstract : The conventional Formalin Fixed Paraffin Embedded (FFPE) technique is the gold standard for preserving histomorphology, while cryopreservation of tissue is the gold standard for biomolecules’ preservation. Once the FFPE tissues stained, the slides are routinely archived along with FFPE blocks for long in biobanks/hospitals and there for underastimated and underused. However, the use of already fixed and stained FFPE biospecimens to extract DNA and/or RNA is not a common practice worldwide. This study provided additional piece of evidence about the feasibility of cover slip removal in less than 24 hours using either Benzene 37 Co or Acetone 37 Co. Following cover slip removal and crude dissection of the immunostained IHC, satisfactory quantity and quality of both DNA and total RNA were extracted and successfully used to perform some downstream molecular analysis. In fact, DNA was used to report higher methylation profiles of HIC1 gene in colorectal carcinoma (CRC) patients. Mutational profiles of selected proto-oncogenes KRAS and CTNNB1 as common mutated candidates in CRC were also studied using this reused DNA. Total RNA with successfeul quality was used for gene expression analysis of RPLPO and miR-124a as a tumor suppressors in CRC. A workflow designed by our group was developed for cover-slip removal by immersion in Acetone, Benzene, Tetrahydrofuran (THF) at different temperatures. Following cover-slip removal and crude dissection of the immunostained IHC, satisfactory quantity and quality of both DNA and total RNA were successfully extracted and successfully used to perform some downstream molecular analysis.This pioneering study suggested a promising approach to recycle and reuse both DNA and RNA from already immunostained slides with suitable concentration and integrity as starting biomolecules/templates for further downstream molecular applications. Such approach if applied at large scale will revolutionize the biobanking approaches worldwide. 
Supervisor : Dr. Salina Yahya Saddick 
Thesis Type : Master Thesis 
Publishing Year : 1440 AH
2019 AD 
Co-Supervisor : Dr. Mourad Taher Assidi 
Added Date : Wednesday, September 11, 2019 

Researchers

Researcher Name (Arabic)Researcher Name (English)Researcher TypeDr GradeEmail
حنين زكي جاويJawi, Haneen ZakiResearcherMaster 

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