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                            Library & Multimedia
 Single Cell Forensic Short Tandem Repeat Typing within Microfluidic Droples
NCJ Number
              255450
Journal

*Analytical Chemistry* Volume: 86 Issue: 1 Dated: 2014 Pages: 703-712
Author(s)
          Tao Geng;
                        Richard Novak;
                        Richard Mathies
Date Published

2014
Length
              10 pages
Annotation
              This article reports on the development of a short tandem repeat (STR) typing method for forensic identification of individual cells.
Abstract
              This method produced monodisperse 1.5 nL agarose-in-oil droplets with a high frequency, using a microfluidic droplet generator. Statistically dilute single cells, along with primer-functionalized microbeads, are randomly compartmentalized in the droplets. Massively parallel single-cell droplet polymerase chain reaction (PCR) is performed to transfer replicas of desired STR targets from the single-cell genomic DNA onto the coencapsulated microbeads. These DNA-conjugated beads are subsequently harvested and reamplified under statistically dilute conditions for conventional capillary electrophoresis (CE) STR fragment size analysis. The 9-plex STR profiles of single cells from both pure and mixed populations of GM09947 and GM09948 human lymphoid cells show that all alleles are correctly called and allelic drop-in/drop-out is not observed. The cell mixture study exhibits a good linear relationship between the observed and input cell ratios in the range of 1:1 to 10:1. Additionally, the STR profile of GM09947 cells could be deduced even in the presence of a high concentration of cell-free contaminating 9948 genomic DNA. This method will be valuable for the STR analysis of samples that contain mixtures of cells/DNA from multiple contributors and for low-concentration samples. Figures, tables, and 36 references (publisher abstract modified)
      
  
          Additional Details
Grant Number(s)
        2009-DN-BX-K180


  
  
        
  Agencies
        NIJ-Sponsored


  
  
        
  Award
        Microchip Analyzer for Forensic Short Tandem Repeat Typing of Single Cells
    
  
  
Date Published: January 1, 2014
    
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