MDA

Multiple Displacement Amplification

MDA is a method commonly used for sequencing microbial genomes due to its ability to amplify templates larger than 0.5 Mbp, but it can also be used to study genomes of other sizes (Dean et al., 2001). In this method, 3ê-blocked random hexamer primers are hybridized to the template, followed by strand-displacement DNA synthesis with Phi 29 polymerase. The method allows for efficient and rapid DNA amplification. Deep sequencing of the amplified DNA provides accurate representation of reads, while sequencing depth provides better alignment and consensus for sequences. Several variations on the original MDA methodãsuch as MIDAS (Gole et al., 2013), ddMDA (Rhee et al., 2016), SNES (Leung et al., 2015), and IMS-MDA (Seth-Smith et al., 2013) ãhave been developed to improve the amplification bias and throughput (Seth-Smith et al., 2013).

Advantages:

  • Templates can be circular DNA (eg, plasmids, bacterial DNA)
  • Can sequence large templates
  • Can perform single-cell sequencing or sequencing for samples with limited amounts of starting material

Disadvantages:


Reagents:

Illumina Library prep and Array Kit Selector



Reviews:

Sauvage V. and Eloit M. Viral metagenomics and blood safety. Transfus Clin Biol. 2016;23:28-38

Zhang X., Marjani S. L., Hu Z., Weissman S. M., Pan X. and Wu S. Single-Cell Sequencing for Precise Cancer Research: Progress and Prospects. Cancer Res. 2016;76:1305-1312

Hou Y., Wu K., Shi X., et al. Comparison of variations detection between whole-genome amplification methods used in single-cell resequencing. Gigascience. 2015;4:37

Wang Y. and Navin N. E. Advances and applications of single-cell sequencing technologies. Mol Cell. 2015;58:598-609



References:

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Troell K., Hallstrom B., Divne A. M., et al. Cryptosporidium as a testbed for single cell genome characterization of unicellular eukaryotes. BMC Genomics. 2016;17:471

Ning L., Li Z., Wang G., et al. Quantitative assessment of single-cell whole genome amplification methods for detecting copy number variation using hippocampal neurons. Sci Rep. 2015;5:11415

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Peterson S. W., Knox N. C., Golding G. R., et al. A Study of the Infant Nasal Microbiome Development over the First Year of Life and in Relation to Their Primary Adult Caregivers Using cpn60 Universal Target (UT) as a Phylogenetic Marker. PLoS One. 2016;11:e0152493

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Leung M. L., Wang Y., Waters J. and Navin N. E. SNES: single nucleus exome sequencing. Genome Biol. 2015;16:55

Li N., Wang L., Wang H., et al. The Performance of Whole Genome Amplification Methods and Next-Generation Sequencing for Pre-Implantation Genetic Diagnosis of Chromosomal Abnormalities. J Genet Genomics. 2015;42:151-159

Macaulay I. C., Haerty W., Kumar P., et al. G&T-seq: parallel sequencing of single-cell genomes and transcriptomes. Nat Methods. 2015;

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Zhang C. Z., Spektor A., Cornils H., et al. Chromothripsis from DNA damage in micronuclei. Nature. 2015;522:179-184