Strategies for low copy number DNA analysis
Keywords
DNA -- Analysis; Forensic genetics -- Technique; Gene amplification; Polymerase chain reaction
Abstract
Low copy number (LCN) DNA analysis is an approach that involves the examination of minute quantities (i.e.,
multiple displacement amplification (MDA). PEP PCR uses a mixture of random 15-base oligonucleotide primers to prime and subsequently amplify the whole genome (or a large percentage of it) prior to subsequent genetic analysis while DOP PCR uses a single degenerate primer. MDA is an isothermal process that employs random hexamer primers to amplify the whole genome. Epithelial and sperm cell suspensions were subjected to PEP PCR, DOP PCR, and MDA. Aliquots of the products were re-amplified using primers specific for the D 1 S80 PCR VNTR locus. Cycle numbers ranged from the standard 30 cycles up to 50 cycles. Results indicated that WGA products from PEP PCR and DOP PCR could be used to analyze <100 pg of DNA and to increase the sensitivity of D1S80 analysis from a few cells to one cell equivalent when the cycle number used for DI S80 analysis was increased above the standard cycle number. Although WGA products from MDA could be used to type a few ( <5) cells, the sensitivity of D 1 S80 analysis was not increased using this WGA method. Allelic drop-out and non-specific amplification were observed with each WGA method tested. The preceding LCN DNA typing strategies used the D1S80 PCR VNTR system as a model. The same strategies are being applied to Y-STR typing systems and the efficacy of these methods for forensic casework specimens is being examined.
Notes
This item is only available in print in the UCF Libraries. If this is your thesis or dissertation, you can help us make it available online for use by researchers around the world by STARS for more information.
Graduation Date
2003
Advisor
Ballantyne, Jack
Degree
Master of Science (M.S.)
College
College of Arts and Sciences
Department
Chemistry
Format
Pages
135 p.
Language
English
Length of Campus-only Access
None
Access Status
Masters Thesis (Open Access)
Identifier
DP0028721
Subjects
Arts and Sciences -- Dissertations, Academic; Dissertations, Academic -- Arts and Sciences
STARS Citation
Raker, Virginia L., "Strategies for low copy number DNA analysis" (2003). Retrospective Theses and Dissertations. 984.
https://stars.library.ucf.edu/rtd/984