![]() ![]() ![]() ![]() Especially for oncology, single-cell sequencing offers novel insights into the evolution of cancers over time and in reaction to treatment, which will lead to novel strategies for treatment regimens and drug development 6. For example, the complexity of alterations of the cancer genome has only been grasped recently by assessing single tumour cells in parallel 3, 4, 5. It has become apparent in the last 5 years that genomic analysis of single cells provides crucial information that is lost in bulk sequencing of tissue because of averaging effects and limitations of computational methods to deconvolute sequence information from many different clones 1, 2. The advantages of this method, which we named TruePrime, promise to facilitate and improve single-cell genomic analysis. Moreover, copy number variant (CNV) calling yields superior results compared with random primer-based MDA methods. This novel method demonstrates superior breadth and evenness of genome coverage, high reproducibility, excellent single-nucleotide variant (SNV) detection rates with low allelic dropout (ADO) and low chimera formation as exemplified by sequencing HEK293 cells. A combination of TthPrimPol’s unique ability to synthesize DNA primers with the highly processive Phi29 DNA polymerase (Φ29DNApol) enables near-complete whole genome amplification from single cells. TthPrimPol displays a potent primase activity preferring dNTPs as substrates unlike conventional primases. Here we introduce a novel multiple displacement amplification (MDA) method based on the unique DNA primase features of Thermus thermophilus ( Tth) PrimPol. Sequencing of a single-cell genome requires DNA amplification, a process prone to introducing bias and errors into the amplified genome. ![]()
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