Single cell DNA amplification
The development of technologies based on single-cell sequencing is rapidly enhancing progress in genomics, epigenetics, transcriptomics and proteomics. The ability to target individual cells will be fundamental in providing a detailed understanding of cell lineage relationships and of the functional states of individual cells.
Oxford researchers have developed a novel method for the highly accurate and reliable amplification of DNA from very low input amounts, particularly from single cells. The Oxford method provides unbiased amplification of nucleic acids. This offers widespread applications as a research tool, and for environmental and clinical diagnostics.
Next generation sequencing
Next Generation Sequencing (NGS) techniques enable entire genomes to be sequenced. NGS technologies have improved precision and throughput whilst becoming increasingly cost effective. A major leap forward has been the increasing ability to perform analyses using limited amounts of material. It is now feasible to analyse single cells thereby offering applications in many scientific fields, including clinical science, which was previously believed to be implausible.
Genomic DNA amplification from individual cells
Existing techniques for the accurate and reliable amplification of nucleic acids from single cells suffer from certain disadvantages. The widely used MDA (Multiple Displacement Amplification) technique, for example, uses random hexamer primers which have a tendency to interact and give rise to template-independent polymerisation. MDA also shows amplification biases, leading to uneven amplification. Alternatively, PCR-based methods suffer from poor genome coverage, GC biases, and allelic preference, where one allele is amplified in preference to another, and allelic dropout, where non-amplification of an allele occurs.
The new method for low input DNA amplification
Oxford researchers have developed a proprietary method for the amplification of nucleic acids using very low amounts of material, which is particularly applicable for single cell analyses. The method is compatible with techniques such as whole-genome sequencing, copy-number profiling, and pre-amplification for targeted analyses such as detection, sequencing or genotyping. The Oxford method confers the following advantages:
- Highly accurate, reliable and even amplification using low amounts of input material
- Unbiased amplification of alleles
- Improved randomness of fragment choice compared to existing methods
- Negligible yield of non-specific products in the absence of source material, thus eliminating a major source of artefacts produced by existing methods
The method has utility for a number of applications, including the study of genomic heterogeneity and other single-cell research, forensics, the study of recombination, analysis of polar bodies, pre-implantation and in utero genetic diagnosis.
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