Date of Defense
4-11-2025 10:00 AM
Location
F3-223
Document Type
Thesis Defense
Degree Name
Master of Science in Molecular Biology and Biotechnology
College
COS
Department
Biology
First Advisor
Dr. Rabah Iratni
Second Advisor
Dr. Rashid Al-Ghafri
Third Advisor
Dr. Sumaya Al-Mansoori
Abstract
DNA degradaon and insufficient gDNA quanty and purity, preserving genomic DNA on FTA Micro-Cards presents substanal difficules for forensic invesgaons of unexpected fatalies. Through the use of lowquality genomic DNA recovered from four-year-old dried blood using FTA Micro-Cards, this effort seeks to enhance and opmize exome sequencing. Samples were sequenced using Next-Generaon Sequencing (NGS) with Illumina DNA Prep with Exome 2.0 Plus Enrichment with mitochondrial panel, starng at 0.1 ng/μl of gDNA.To improve the coverage of mitochondrial genes, a mitochondrial DNA spike-in was included, and the samples were sequenced using the NextSeq 1000/2000 plaorm, employing the onboard DRAGEN Enrichment Pipeline for analysis.
Unusual A260/A280 and A260/A230 purity raos were noted during the quality control (QC) of the extracted gDNA, indicang that the gDNA amounts were inadequate and failed to fulfill the specified standards for exome methods. Notwithstanding these drawbacks, the research employed 16 plex methodology to proficiently augment sequencing using modified protocols for library preparaon and enrichment. High coverage through enre exome sequencing at depths of 30X and above revealed that dependable genomic data may be acquired from low-quality inputs from samples with restricted recollectability.
These results highlight the ability to extract valuable genec informaon from contaminated materials, parcularly for forensic purposes and the detecon of genec variaons linked to unexpected mortality. The importance of older FTA samples is increased by this work, which also highlights the need for specialized opmizaon methods when handling biological specimens that have been degraded.
These results highlight the ability to discover genec variaons linked to sudden death and to retrieve crucial genec informaon from degraded materials, especially in forensic circumstances. This study enhances the suitability of legacy FTA Micro-Cards in complex forensic invesgaons and emphasizes the significance of customized opmizaon techniques for handling deteriorated biological specimens. This study involves the opmizaon of aged FTA cards in forensic challenging invesgaons, highlighng the need of customized improved techniques for the management of challenging biological specimens.
Included in
ADAPTATION OF EXOME SEQUENCING METHODS FOR LOW-QUALITY GENOMIC DNA EXTRACTED FROM AGED FTA MICRO-CARDS IN SUDDEN DEATH INVESTIGATIONS
F3-223
DNA degradaon and insufficient gDNA quanty and purity, preserving genomic DNA on FTA Micro-Cards presents substanal difficules for forensic invesgaons of unexpected fatalies. Through the use of lowquality genomic DNA recovered from four-year-old dried blood using FTA Micro-Cards, this effort seeks to enhance and opmize exome sequencing. Samples were sequenced using Next-Generaon Sequencing (NGS) with Illumina DNA Prep with Exome 2.0 Plus Enrichment with mitochondrial panel, starng at 0.1 ng/μl of gDNA.To improve the coverage of mitochondrial genes, a mitochondrial DNA spike-in was included, and the samples were sequenced using the NextSeq 1000/2000 plaorm, employing the onboard DRAGEN Enrichment Pipeline for analysis.
Unusual A260/A280 and A260/A230 purity raos were noted during the quality control (QC) of the extracted gDNA, indicang that the gDNA amounts were inadequate and failed to fulfill the specified standards for exome methods. Notwithstanding these drawbacks, the research employed 16 plex methodology to proficiently augment sequencing using modified protocols for library preparaon and enrichment. High coverage through enre exome sequencing at depths of 30X and above revealed that dependable genomic data may be acquired from low-quality inputs from samples with restricted recollectability.
These results highlight the ability to extract valuable genec informaon from contaminated materials, parcularly for forensic purposes and the detecon of genec variaons linked to unexpected mortality. The importance of older FTA samples is increased by this work, which also highlights the need for specialized opmizaon methods when handling biological specimens that have been degraded.
These results highlight the ability to discover genec variaons linked to sudden death and to retrieve crucial genec informaon from degraded materials, especially in forensic circumstances. This study enhances the suitability of legacy FTA Micro-Cards in complex forensic invesgaons and emphasizes the significance of customized opmizaon techniques for handling deteriorated biological specimens. This study involves the opmizaon of aged FTA cards in forensic challenging invesgaons, highlighng the need of customized improved techniques for the management of challenging biological specimens.