Knowledge Resource Center for Ecological Environment in Arid Area
DOI | 10.2144/000114389 |
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
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ISSN | 0736-6205 |
EISSN | 1940-9818 |
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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