Related Experiment Video
Updated: Apr 10, 2026
09:45
Detection of Copy Number Alterations Using Single Cell Sequencing
Published on: February 17, 2017
12.3K
Single-Cell Whole-Genome Amplification and Sequencing: Methodology and Applications
Lei Huang1, Fei Ma, Alec Chapman
1Biodynamic Optical Imaging Center (BIOPIC), School of Life Sciences, Peking University, Beijing 100871, China.
Annual Review of Genomics and Human Genetics
|June 17, 2015
Summary
This study evaluates single-cell whole-genome amplification (WGA) methods like MDA and MALBAC. It defines key performance metrics to compare WGA kits for applications in cancer genomics and genetic diagnostics.
Area of Science:
- Genomics
- Molecular Biology
Background:
- Single-cell whole-genome amplification (WGA) is crucial for genomic analysis of individual cells.
- Various WGA methods exist, each with distinct performance characteristics.
Purpose of the Study:
- To survey and define performance parameters for single-cell WGA methods.
- To compare commercial WGA kits using deep sequencing.
- To discuss applications of single-cell genomics.
Main Methods:
- Evaluated degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR), multiple displacement amplification (MDA), and multiple annealing and looping-based amplification cycles (MALBAC).
- Defined key performance metrics: genome coverage, uniformity, reproducibility, unmappable rates, chimera rates, allele dropout, false positive rates for single-nucleotide variations, and copy-number variation detection.
- Performed deep sequencing on multiple single cells from five commercial WGA kits.
Main Results:
- Commercial WGA kits were compared based on defined performance metrics.
- Detailed characterization of WGA method performance was achieved through deep sequencing.
Conclusions:
- The study provides a framework for evaluating and selecting appropriate WGA methods for single-cell genomics.
- Identified key parameters and compared commercial kits, aiding researchers in applications like cancer genomics and preimplantation genetic diagnosis.
Related Concept Videos
03:21RNA-seq
12.6K
RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases.
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
12.6K
03:00Next-generation Sequencing
101.8K
The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features....
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features....
101.8K
01:57Sanger Sequencing
779.8K
DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...
779.8K