U.S. flag
      An official website of the United States government, Department of Justice.
        Here's how you know
      Here's how you know**Official websites use .gov**
            
            A **.gov** website belongs to an official government organization in the United States.
      **Secure .gov websites use HTTPS**
            
            A **lock** (
            A locked padlock
            ) or **https://** means you’ve safely connected to the .gov website. Share sensitive information only on official, secure websites. Menu

      
        close

            
            
                                              About NIJ
        
                    
  
          About NIJ
      
    
          The NIJ Director
          How NIJ Is Organized
          Mission and Guiding Principles
          Research and Development Process
          Strategic Framework
          Assessments of NIJ
          Annual Reports & Reports to Congress
          Staff Directory
          Funding & Awards
        
                    
  
          Funding & Awards
      
    
          Current Funding
          Forthcoming Funding
          Expired Funding
          Awards: Listing of Funded Projects
          Guidance for Applicants and Awardees
          Fellowship & Student Programs
          Travel Scholarships
          NIJ Challenge Program
          Library & Multimedia
        
                    
  
          Library & Multimedia
      
    
          NIJ Journal
          Podcasts
          Publications Listing
          Multimedia Listing
          Rated Practices on CrimeSolutions
          Rated Programs on CrimeSolutions
          Software & Databases
          Datasets
          Patents
          Topics
        
                    
  
          Topics
      
    
          Article Listing
          Corrections
          Courts
          Crime Prevention
          Crimes
          Drugs and Crime
          Equipment and Technology
          Forensic Sciences
          Youth Justice
          Justice System Reform
          Law Enforcement
          Tribal Crime and Justice
          Victims of Crime
          Events
        
                    
  
          Events
      
    
          NIJ 2024 Research Conference
          Scheduled Events
          On-Demand Events
          Research Meetings & Workshops
          NIJ-Hosted Online Training

                            
                                About
          Contact Us
          Subscribe
          Topics A-Z
                  Share
FacebookXLinkedInRedditPinterestEmail Home
                            Library & Multimedia
 Novel Thermostable Y-Family Polymerases: Applications for the PCR Amplification of Damaged or Ancient DNAs
NCJ Number
              241189
Journal

*Nucleic Acids Research* Volume: 34 Issue: 4 Dated: 2006 Pages: 1102-1111
Author(s)
          John P. McDonald;
                        Ashley Hall;
                        Didier Gasparutto;
                        Jean Cadet;
                        Jack Ballantyne;
                        Roger Woodgate
Date Published

2006
Length
              9 pages
Annotation
              The research reported in this article provides "proof of principle" that the inclusion of a thermostable Y-family DNA polymerase in the PCR amplification of damaged DNA facilitates the recovery of amplicons previously unobtainable with Taq polymerase alone.
Abstract
              For many years, Taq polymerase has served as the stalwart enzyme in the PCR amplification of DNA; however, a major limitation of Taq is its inability to simplify damaged DNA, thereby restricting its usefulness in forensic applications. This article reports the identification and characterization of five novel thermostable Dpo4-like enzymes from Acidianus infernus, Sulfolobus shibatae, Sulfolobus tengchongensis, Stygioiobus azoricus, and Sulfurisphaera ohwkuensis, as well as two recombinant chimeras that have enhanced enzymatic properties compared with the naturally occurring polymerases. By using a blend of Taq and Dpo4-like enzymes, the authors obtained a PCR amplicon from ultraviolet-irradiated DNA that was largely unamplifyable with Taq alone. This research involves the first steps toward using a Y-family polymerase to replicate the damaged DNA template, so as to generate one or more lesion-free copies of the template that could be subsequently amplified by a higher fidelity PCR enzyme. Descriptions of materials and methods address the identification and cloning of novel Dpo4-like genes, the construction of chimeric enzymes, the purification of Dpo4-like enzymes, in vitro primer extension assays, PCRs on undamaged DNA, Genomic DNA sample preparation, and PCR amplification and detection of Alu sequences. 6 figures and 28 references
      
  
          Additional Details
Grant Number(s)
        2003-IJ-CX-K001
                                MRTN-CT 2003-505086


  
  
        
  Agencies
        NIJ-Sponsored


  
  
        
  Publication Type
        Report (Study/Research)


  
  
        
  Award
        National Institute of Justice Congressionally Directed Awards
    
  
  
Date Published: January 1, 2006
    
          Downloads
        
                      HTML

Side navigation
  

        

  
  
              NIJ Journal
    
      
            
            
            
      Podcasts
    
      
            
            
            
      Publications Listing
    
      
            
            
            
      Multimedia Listing
    
      
            
            
            
      Rated Practices on CrimeSolutions
    
      
            
            
            
      Rated Programs on CrimeSolutions
    
      
            
            
            
      Software & Databases
    
      
            
            
            
      Datasets
    
      
            
            
            
      PatentsRelated Topics
        
                DNA fingerprinting
        Forensic sciences
        Investigative techniques
        NIJ Grant-related Documents
        Suspect identification



  
Similar Publications
        
                      Near-infrared Excitation Raman Analysis of Underlying Colorants on Redyed Fabric
                  Moving Forward While Looking Back: Understanding the Influence of a “Test All” SAK Mandate on Sexual Assault Case Attrition
                  Assessing Physical Fit Examinations of Stabbed and Torn Textiles Through a Large Dataset of Casework-like Items and Interlaboratory Studies
 Top

      National Institute of Justice

        
        
                        About NIJ
                    About NIJ
      
    
                  How NIJ Is Organized
      
    
                  NIJ Director
      
    
                  Research Disclaimer
      
    
                  Funding & Awards
                    Funding & Awards Home
      
    
                  Awards Listing
      
    
                  Current Funding
      
    
                  Guidance for Applicants and Awardees
      
    
                  Resources
                    Events
      
    
                  Podcasts
      
    
                  Publications
      
    
                  Topics
      
    
                  Multimedia
      
    
                  Reuse Policy
      
    
                  Contacts
                    Contact NIJ
      
    
                  Staff Directory
      
    
                  Submit a FOIA Request
      
    
                  File a Civil Rights ComplaintFacebook
          
                      X
          
                      YouTube
  

      810 7th St. NW, Washington, D.C. 20531
      
              Secondary Footer link menu
            Accessibility
              Plain Language
              Privacy Policy
              Legal Policies and Disclaimer
              No FEAR Act
              Freedom of Information Act
              USA.gov
              Justice.gov