Yikon Genomics

Yikon Genomics is a leading solution-based single cell sequencing company. Currently based on the patented MALBAC^TM technology from Harvard University, Yikon provides complete solutions in the fields of reproductive health and cancer diagnostics with liquid tumor biopsy. The MALBAC-PGSTM and MALBAC-PGDTM solutions offer the most accurate and comprehensive embryo testing results for physicians to choose the best embryos with balanced chromosomal copy numbers,without inheriting the known disease-associated alleles. The MALBAC-PGSTM and MALBAC-PGDTM solutions have helped thousands of couples to achieve successful pregnancies. Yikon will continue to interpret the code of life and bring people the hope and joy of life.

Overview

Based on the patented MALBAC™ technology from Harvard University, MALBAC™ Single Cell WGA kit provides rapid, reliable and reproducible whole genome amplification of single cell(s) or picogram to sub-nanogram quantities of gDNA to produce 2-4 micrograms of amplified products in about 4 hours.

Principle

Nucleic acid analytical techniques such as real-time qPCR, microarray and next-generation sequencing often require starting nucleic acid meet specific quantity and quality threshold to generate reproducible and accurate data. Therefore, Whole genome amplification (WGA) is often required prior to analyze samples with limited quantity. Such a typical necessity is single cell sequencing. Single cells WGA followed by genome sequencing is highly desirable for many applications such as pre-implantation genetic screening/diagnosis regarding in vitro fertilization, or genotyping of rare circulation tumor cells.

WGA methods are in general prone to amplification bias. Conventional PCR-based WGA methods such as Degenerate Oligonucleotide Primed PCR (DOP-PCR) and Primer Extension Pre-amplification (PEP) innately introduce sequence-dependent biases, and the biases are magnified accumulatively over each round of thermal cycling during the exponential amplification. Multiple Displacement Amplification (MDA) method, in which random primer and the strand-displacing phi29 polymerase are used under isothermal condition, improves amplification evenness over the conventional PCR-based methods by avoiding thermal cycling. However, as the entire amplification process of MDA is driven by random hybridization of hexamer, the primer degeneracy caused biases accumulation during the amplification, thus MDA method still exhibits considerable biases in its amplicons.

MALBAC^TM (Multiple Annealing and Looping Based Amplification Cycles) technology, which carries out a few quasilinear pre-amplification cycles first bytailed random primer, then the amplification enters exponential phase driven by universal primer (the tail sequence) until micrograms PCR products are generated. The amplification by universal primer minimizes bias generation in amplicons thus the uniformity of MALBAC^TM is far better than any other WGA method up to date.

Advantages

Generally, there are 8 key parameters to characterize the performance of whole genome amplification methods, including genome coverage, uniformity, reproducibility, unmappable rates, chimera rates, allele dropout rates, false positive rates for calling single-nucleotide variations, and ability to call copy-number variations. MALBAC performs better among other whole genome amplification methods.

– Amplify single cells genome to ug level, and only need about 2-4 hrs in one single tube.

– Best amplification uniformity among other similar products, Single-Nucleotide Mismatch rate～10^-5

– High Coverage that >90% locus can be successfully amplified, locus allele drop out <10%

– Various Applications-The amplified products can be applied for copy number variation (CNV) and single nucleotide polymorphism (SNP) analysis, pre-implantation genetic screening (PGS) and diagnosis (PGD), circulating tumor cell (CTC), etc.

Coefficient of variation (CV) is the key parameter for copy-number variation (CNV) analysis by next-generation sequencing, which is important for accurate measurements of CNV; Yikon’s MALBAC Single Cell WGA Kit is superior to other commercial kits regarding CV parameter with more even CNV data after normalization. This over advantage makes MALBAC Single Cell WGA Kit more suitable for clinical application, in particular for preimplantation genetic screening (PGS).

The allele dropout rate (ADO) is one of the most characteristics of whole genome amplification, particularly for medical application such as preimplantation genetic diagnosis (PGD). ADO is also the primary cause of false negatives of SNV calling (point mutation). MALBAC Single Cell WGA Kit has lower allele dropout rates than the other commercial kits, which indicates MALBAC Single Cell WGA Kit is the first choice for PGD and SNV analysis.

Applications

Mammalian Animal:

 - Pre-implantation genetic screening

 - Pre-implantation genetic diagnosis

 - Genotyping of transgenic animals

 - Phasing of individual human genomes

 - Genotyping of sperm cell

 - Embryo and stem cell

 - Forensic specimens analysis

 - Typing of transgenic animals

Tumor:

 - Somatic genetic variation analysis

 - Tumor evolution and development

 - Tumor stem cell

 - Circulating tumor cells

Bacteria:

 - Genotyping of microorganism

 - Metagenomics study

 - Pathogen analysis

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