Abstract
To allow multiple genetic analyses on a single cell, whole genome amplification (WGA) is required. Unfortunately, studies comparing different WGA methods for downstream human identification Short Tandem Repeat (STR) analysis remain absent. Therefore, the aim of this work was to assess the performance of four commercially available WGA kits for downstream human identification STR profiling on a B-lymphoblastoid cell line. The performance was assessed using an input of one or three micromanipulated cells. REPLI-g showed a very low dropout rate, as it was the only WGA method in this study that could provide a complete STR profile in some of its samples. Although Ampli1, DOPlify and PicoPLEX did not detect all selected STR markers, they seem suitable for genetic identification in single-cell applications.
Highlights
Whole genome amplification (WGA) is inevitable to allow multiple genetic analyses on either a single cell or a limited number of cells
Previous research by Denis et al demonstrates that multiple displacement amplification (MDA) whole genome amplification (WGA) might be applicable for Short Tandem Repeat (STR) analysis[10]
REPLI-g Single Cell Kit, PicoPLEX WGA Kit, Ampli[1] WGA Kit and DOPlify WGA were compared in parallel for their ability to produce WGA product, adequate for human identification STR analysis
Summary
Whole genome amplification (WGA) is inevitable to allow multiple genetic analyses on either a single cell or a limited number of cells. The growing interest in single-cell analysis for clinical applications raises the need to identify single cells Such a genetic analysis that is often performed on only a limited number of cells is STR analysis for human identification DNA profiling. One single highly specific primer, complementary to the adapters that are ligated on both sides of each DNA fragment, initiates the subsequent PCR amplification reaction This ligation-based WGA method has already been successfully applied in a cbNIPT context[11]. DOPlify Reaction Kit is an advanced version of this classical DOP-PCR and has recently shown promising results for CNV analysis, with minimal bias introduction[6] This kit has not yet been tested in the context of STR profiling. A bulk DNA sample from the B-LCL was included in the STR-PCR reaction and used as a reference STR-profile against which all sample results were compared
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