Arid
DOI10.2144/000114389
Improved lysis of single bacterial cells by a modified alkaline-thermal shock procedure
He, Jian1,2; Du, Shiyu3; Tan, Xiaohua4; Arefin, Ayesha2; Han, Cliff S.2
通讯作者He, Jian ; Han, Cliff S.
来源期刊BIOTECHNIQUES
ISSN0736-6205
EISSN1940-9818
出版年2016
卷号60期号:3页码:129-135
英文摘要

Single-cell genomics (SCG) is a recently developed tool to study the genomes of unculturable bacterial species. SCG relies on multiple-strand displacement amplification (MDA), PCR, and next-generation sequencing (NGS); however, obtaining sufficient amounts of high-quality DNA from samples is a major challenge when performing this technique. Here we present an improved bacterial cell lysing procedure that combines incubation in an alkaline buffer with a thermal shock (freezing/heating) treatment to yield highly intact genomic DNA with high efficiency. This procedure is more efficient in lysing Bacillus subtilis and Synechocystis cells compared with two other frequently used lysis methods. Furthermore, 16S ribosomal RNA gene and overall genome recovery were found to be improved by this method using single cells from a Utah desert soil community or Escherichia coli single cells, respectively. The efficiency of genome recovery for E. coli single cells using our procedure is comparable with that of the REPLI-g Single Cell (SC) Kit, but our method is much more economical. By providing high-quality genome templates suitable for downstream applications, our procedure will be a promising improvement for SCG research.


英文关键词cell lysis single-cell genomics genome recovery unculturable microbiology genome sequencing
类型Article
语种英语
国家Peoples R China ; USA
收录类别SCI-E
WOS记录号WOS:000372102200005
WOS关键词SP-NOV. ; AMPLIFICATION ; DIVERSITY ; RESISTANCE ; GENOMICS ; SAMPLES ; SOIL
WOS类目Biochemical Research Methods ; Biochemistry & Molecular Biology
WOS研究方向Biochemistry & Molecular Biology
资源类型期刊论文
条目标识符http://119.78.100.177/qdio/handle/2XILL650/191830
作者单位1.Chinese Acad Sci, Dalian Inst Chem Phys, Ctr Translat Med, Dept Biotechnol, Dalian, Liaoning, Peoples R China;
2.Los Alamos Natl Lab, Biosci Div, Los Alamos, NM USA;
3.Chinese Acad Sci, Engn Lab Specialty Fibers & Nucl Energy Mat, Ningbo Inst Mat Technol & Engn, Ningbo, Zhejiang, Peoples R China;
4.Wenzhou Med Univ, Key Lab Biotechnol & Pharmaceut Engn, Mol Pharmacol Res Ctr, Sch Pharm, Wenzhou, Zhejiang, Peoples R China
推荐引用方式
GB/T 7714
He, Jian,Du, Shiyu,Tan, Xiaohua,et al. Improved lysis of single bacterial cells by a modified alkaline-thermal shock procedure[J],2016,60(3):129-135.
APA He, Jian,Du, Shiyu,Tan, Xiaohua,Arefin, Ayesha,&Han, Cliff S..(2016).Improved lysis of single bacterial cells by a modified alkaline-thermal shock procedure.BIOTECHNIQUES,60(3),129-135.
MLA He, Jian,et al."Improved lysis of single bacterial cells by a modified alkaline-thermal shock procedure".BIOTECHNIQUES 60.3(2016):129-135.
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