Topic
MALBAC
About: MALBAC is a research topic. Over the lifetime, 113 publications have been published within this topic receiving 9367 citations. The topic is also known as: MALBAC.
Papers published on a yearly basis
Papers
TL;DR: MDA-based whole genome amplification by multiple displacement amplification is a simple and reliable method that could have significant implications for genetic studies, forensics, diagnostics, and long-term sample storage.
Abstract: Fundamental to most genetic analysis is availability of genomic DNA of adequate quality and quantity. Because DNA yield from human samples is frequently limiting, much effort has been invested in developing methods for whole genome amplification (WGA) by random or degenerate oligonucleotide-primed PCR. However, existing WGA methods like degenerate oligonucleotide-primed PCR suffer from incomplete coverage and inadequate average DNA size. We describe a method, termed multiple displacement amplification (MDA), which provides a highly uniform representation across the genome. Amplification bias among eight chromosomal loci was less than 3-fold in contrast to 4–6 orders of magnitude for PCR-based WGA methods. Average product length was >10 kb. MDA is an isothermal, strand-displacing amplification yielding about 20–30 μg product from as few as 1–10 copies of human genomic DNA. Amplification can be carried out directly from biological samples including crude whole blood and tissue culture cells. MDA-amplified human DNA is useful for several common methods of genetic analysis, including genotyping of single nucleotide polymorphisms, chromosome painting, Southern blotting and restriction fragment length polymorphism analysis, subcloning, and DNA sequencing. MDA-based WGA is a simple and reliable method that could have significant implications for genetic studies, forensics, diagnostics, and long-term sample storage.
1,694 citations
TL;DR: DOP-PCR represents a rapid, efficient, and species-independent technique for general DNA amplification, and has advantages over interspersed repetitive sequence PCR (IRS- PCR), which relies on the appropriate positioning of species-specific repeat elements.
1,573 citations
TL;DR: A new amplification method—multiple annealing and looping-based amplification cycles (MALBAC)—that offers high uniformity across the genome that achieves 93% genome coverage ≥1x for a single human cell at 25x mean sequencing depth is reported.
Abstract: Kindred cells can have different genomes because of dynamic changes in DNA. Single-cell sequencing is needed to characterize these genomic differences but has been hindered by whole-genome amplification bias, resulting in low genome coverage. Here, we report on a new amplification method-multiple annealing and looping-based amplification cycles (MALBAC)-that offers high uniformity across the genome. Sequencing MALBAC-amplified DNA achieves 93% genome coverage ≥1x for a single human cell at 25x mean sequencing depth. We detected digitized copy-number variations (CNVs) of a single cancer cell. By sequencing three kindred cells, we were able to identify individual single-nucleotide variations (SNVs), with no false positives detected. We directly measured the genome-wide mutation rate of a cancer cell line and found that purine-pyrimidine exchanges occurred unusually frequently among the newly acquired SNVs.
1,205 citations
TL;DR: W Whole genome amplification beginning with a single cell, or other samples with very small amounts of DNA, has significant implications for multipoint mapping by sperm or oocyte typing and possibly for genetic disease diagnosis, forensics, and the analysis of ancient DNA samples.
Abstract: We have developed an in vitro method for amplifying a large fraction of the DNA sequences present in a single haploid cell by repeated primer extensions using a mixture of 15-base random oligonucleotides. We studied 12 genetic loci and estimate that the probability of amplifying any sequence in the genome to a minimum of 30 copies is not less than 0.78 (95% confidence). Whole genome amplification beginning with a single cell, or other samples with very small amounts of DNA, has significant implications for multipoint mapping by sperm or oocyte typing and possibly for genetic disease diagnosis, forensics, and the analysis of ancient DNA samples.
1,065 citations
TL;DR: A high-throughput method for single-cell whole-genome analysis that was used to measure the genomic diversity in one individual's gamete genomes and found evidence for meiotic drive and evidence for gene conversion.
562 citations
Related Topics (5)
Performance Metrics
| Year | Papers |
|---|---|
| 2022 | 1 |
| 2021 | 4 |
| 2020 | 7 |
| 2019 | 7 |
| 2018 | 6 |
| 2017 | 8 |