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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)

Date Published: January 1, 2014