NDLI: An evaluation of multiple annealing and looping based genome amplification using a synthetic bacterial community

Content Provider	Springer Nature Link
Author	Wang, Yong Gao, Zhaoming Xu, Ying Li, Guangyu He, Lisheng Qian, Peiyuan
Copyright Year	2016
Abstract	The low biomass in environmental samples is a major challenge for microbial metagenomic studies. The amplification of a genomic DNA was frequently applied to meeting the minimum requirement of the DNA for a high-throughput next-generation-sequencing technology. Using a synthetic bacterial community, the amplification efficiency of the Multiple Annealing and Looping Based Amplification Cycles (MALBAC) kit that is originally developed to amplify the single-cell genomic DNA of mammalian organisms is examined. The DNA template of 10 pg in each reaction of the MALBAC amplification may generate enough DNA for Illumina sequencing. Using 10 pg and 100 pg templates for each reaction set, the MALBAC kit shows a stable and homogeneous amplification as indicated by the highly consistent coverage of the reads from the two amplified samples on the contigs assembled by the original unamplified sample. Although GenomePlex whole genome amplification kit allows one to generate enough DNA using 100 pg of template in each reaction, the minority of the mixed bacterial species is not linearly amplified. For both of the kits, the GC-rich regions of the genomic DNA are not efficiently amplified as suggested by the low coverage of the contigs with the high GC content. The high efficiency of the MALBAC kit is supported for the amplification of environmental microbial DNA samples, and the concerns on its application are also raised to bacterial species with the high GC content.
Starting Page	131
Ending Page	136
Page Count	6
File Format	PDF
ISSN	0253505X
Journal	Acta Oceanologica Sinica
Volume Number	35
Issue Number	2
e-ISSN	18691099
Language	English
Publisher	The Chinese Society of Oceanography
Publisher Date	2016-02-24
Publisher Place	Beijing
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	bacterial DNA MALBAC metagenome amplification Oceanography Climatology Ecology Engineering Fluid Dynamics Marine & Freshwater Sciences Environmental Chemistry
Content Type	Text
Resource Type	Article
Subject	Aquatic Science Oceanography

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