It is now possible to amplify DNA to yield a highly reproducible library from a single cell: PicoPLEX whole genome amplification (WGA) technology is designed and optimized for amplification of single copy genomic DNA starting with a single cell. The easy-to-use single tube protocol reduces handling errors, dramatically improves time to results and reduces background. The PicoPLEX WGA Kit can also be used with isolated gDNA amounts ranging from less than 6 pg to 50 pg. The workflow has been successfully used in a broad range of applications for qPCR, arrays, and next-generation sequencing.

A three-step, single-tube reaction starts with lysis of 1–15 cells. The DNA is then used as a template for pre-amplification using a quasi-random priming approach that creates a library of hairpin molecules. These hairpins, in turn, are directly amplified into bulk quantities of DNA that can be used for a wide array of downstream applications.

Flow-sorted cancer cells were amplified with PicoPLEX single cell WGA technology. Each of the cells (blue lines) amplified at the same rate and resulted in a similar, predictable yield. A pooled sample of five cells (red line) represents a positive control. Samples containing 0 cells (green lines; no template control) show very low background.

Single-blastomere biopsies were amplified using PicoPLEX single cell WGA technology, labeled, and hybridized to BlueGnome's 24Sure arrays at Genesis Genetics Institute. Note the clear indication of CNVs. In 2011, ESHRE clinical trials confirmed accuracy of karyotyping using PicoPLEX technology.

Locus-specific qPCR was used to quantify 48 loci in independent single-cell libraries. Data shown compares results of DNA from two individual samples amplified with (left to right) PicoPLEX single cell WGA technology, GenomePLEX, and GenomiPhi using 10 pg of DNA. More than 90% of the product from the PicoPLEX single cell WGA technology produced identifiable, highly reproducible human sequences.