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SIMPLIFIED LOW COPY NUMBER DNA ANALYSIS BY POST PCR PURIFICATION

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Date Issued:
2006
Abstract/Description:
Frequently evidentiary items contain an insufficient quantity of DNA to obtain complete or even partial DNA profiles using standard forensic gentotyping techniques. Here, various methods of post PCR purification were evaluated for their effects on the sensitivity of fluophore-based allelic detection. A method of post PCR purification is described which increases the sensitivity of standard 28 cycle PCR such that low copy number DNA templates (<100 pg DNA) can be analyzed. Full profiles were consistently obtained with as little as 20 pg template DNA without increased cycle number. In mock case type samples with dermal ridge fingerprints, genetic profiles were obtained by amplification with 28 cycles followed by post-PCR purification whereas no profiles were obtained without purification of the PCR product. Allele drop-out, increased stutter, and contamination (allele drop-in) typical of LCN analysis were observed. A single incident of contamination was observed in a reagent blank (not duplicated upon re-amplification) however, no contamination was observed in negative amplification controls.
Title: SIMPLIFIED LOW COPY NUMBER DNA ANALYSIS BY POST PCR PURIFICATION.
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Name(s): Smith, Pamela, Author
Ballantyne, Jack, Committee Chair
University of Central Florida, Degree Grantor
Type of Resource: text
Date Issued: 2006
Publisher: University of Central Florida
Language(s): English
Abstract/Description: Frequently evidentiary items contain an insufficient quantity of DNA to obtain complete or even partial DNA profiles using standard forensic gentotyping techniques. Here, various methods of post PCR purification were evaluated for their effects on the sensitivity of fluophore-based allelic detection. A method of post PCR purification is described which increases the sensitivity of standard 28 cycle PCR such that low copy number DNA templates (<100 pg DNA) can be analyzed. Full profiles were consistently obtained with as little as 20 pg template DNA without increased cycle number. In mock case type samples with dermal ridge fingerprints, genetic profiles were obtained by amplification with 28 cycles followed by post-PCR purification whereas no profiles were obtained without purification of the PCR product. Allele drop-out, increased stutter, and contamination (allele drop-in) typical of LCN analysis were observed. A single incident of contamination was observed in a reagent blank (not duplicated upon re-amplification) however, no contamination was observed in negative amplification controls.
Identifier: CFE0001003 (IID), ucf:46841 (fedora)
Note(s): 2006-05-01
M.S.
Arts and Sciences, Department of Chemistry
Masters
This record was generated from author submitted information.
Subject(s): post PCR purification
low copy number
LCN
MinElute
Microcon-50
Montage PCR
Exo-SAP-IT
Persistent Link to This Record: http://purl.flvc.org/ucf/fd/CFE0001003
Restrictions on Access: public
Host Institution: UCF

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