Assessment of sequence variation within commonly encountered human alpha fibrinogen (HumFGA) alleles

Keywords

DNA -- Analysis, Human genetics -- Variation, Nucleotide sequence

Abstract

The primary goal ofthis !esearch project was to determine if sequence variation exists between individuals who possess the same sized HumFGA alleles. A comprehensive set of procedures was developed that were used to isolate the DNA from blood samples, locate the HumFGA allele of interest and subsequently sequence the DNA fragments and determine if sequence variation existed in the flanking regions and/or the repeat region ( e.g., TTTC) for the HumFGA loci. Blood samples were collected from 104 convicted offenders from the Virginia Division of Forensic Science's DNA Data Bank who possessed a HumFGA allele 21, 22, 23, or 24. The samples collected for this study were obtained from individuals from the Caucasian, African American, and Hispanic population groups in order to determine if sequence variation might be observed more predominantly in a particular population group. To eliminate potential errors due to base misincorporation during the sequencing process both the forward and reverse strands ofDNA fragments were sequenced. In addition, to ensure that any sequence variation observed was genuine and not due to base misincorporation, several colonies from a single sample were sequenced. To further examine the possibility of detecting sequence variations at the HumFGA locus, sizing data for all ofthe samples sequenced, including additional samples, totaling 619 samples containing the HumFGA alleles 21 (196), 22 (129), 23 (121), and 24 (173) were examined to look for any sizing trends that may occur. The sizing data collected from 27 polyacrylamide typing gels containing ~amples amplified using the GenePrint® PowerPlexTM 2.1 System Kit (Promega Corporation, Madison, WI) and analyzed on a FMBIO II Fluorescent Image Analysis System (Hitachi Genetic Systems/MiriaBio, Alameda, CA) were examined. The sizing data from samples at a particular allele (i.e., 21), which were analyzed on the same typing gel, were compared across all 27 polyacrylamide typing gels (Tables 3-5 through 3-8). To further evaluate any structural differences that may exist at the HumFGA 21, 22, 23, or 24 alleles, which might cause the DNA fragments at this locus to migrate slightly differently on a polyacrylamide typing gel than other commonly used forensic loci, the HumFGA sizing data were compared to sizing data obtained for the Hum.CSFlPO 11, 12, and 13 alleles. No sequence variation was found in the flanking or repeat regions for any of the samples sequenced. Therefore, if sequence variation for the HumFGA locus does exist, it is rare and will not routinely be observed.

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

2002

Advisor

Ballantyne, Jack

Degree

Master of Science (M.S.)

College

College of Arts and Sciences

Department

Chemistry

Format

PDF

Pages

124

Language

English

Length of Campus-only Access

None

Access Status

Masters Thesis (Open Access)

Identifier

DP0028720

Subjects

Arts and Sciences -- Dissertations, Academic; Dissertations, Academic -- Arts and Sciences

Accessibility Status

PDF accessibility verified using Adobe Acrobat Pro accessibility checker.

This document is currently not available here.

Share

COinS