High Efficiency Hydrodynamic DNA Fragmentation in a Bubbling System
Li, LH; Jin, ML; Sun, CL; Wang, XX; Xie, ST; Zhou, GF; Van den Berg, A; Eijkel, JCT; Shui, LL; shuill@m.scnu.edu.cn
2017
发表期刊SCIENTIFIC REPORTS
卷号7页码:40745-
摘要DNA fragmentation down to a precise fragment size is important for biomedical applications, disease determination, gene therapy and shotgun sequencing. In this work, a cheap, easy to operate and high efficiency DNA fragmentation method is demonstrated based on hydrodynamic shearing in a bubbling system. We expect that hydrodynamic forces generated during the bubbling process shear the DNA molecules, extending and breaking them at the points where shearing forces are larger than the strength of the phosphate backbone. Factors of applied pressure, bubbling time and temperature have been investigated. Genomic DNA could be fragmented down to controllable 1-10 Kbp fragment lengths with a yield of 75.30-91.60%. We demonstrate that the ends of the genomic DNAs generated from hydrodynamic shearing can be ligated by T4 ligase and the fragmented DNAs can be used as templates for polymerase chain reaction. Therefore, in the bubbling system, DNAs could be hydrodynamically sheared to achieve smaller pieces in dsDNAs available for further processes. It could potentially serve as a DNA sample pretreatment technique in the future.
部门归属[Li, Lanhui; Jin, Mingliang; Xie, Shuting; Zhou, Guofu; Shui, Lingling] South China Normal Univ, Guangzhou Higher Educ Mega Ctr, South China Acad Adv Optoelect, Inst Elect Paper Displays, Guangzhou 510006, Guangdong, Peoples R China; [Li, Lanhui; Jin, Mingliang; Xie, Shuting; Zhou, Guofu; Van den Berg, Albert; Eijkel, Jan C. T.; Shui, Lingling] South China Normal Univ, Guangzhou Higher Educ Mega Ctr, Chinese Minist Educ, Joint Int Res Lab Opt Informat, Guangzhou 510006, Guangdong, Peoples R China; [Sun, Chenglong; Wang, Xiaoxue] Chinese Acad Sci, South China Sea Inst Oceanol, Guangzhou 510301, Guangdong, Peoples R China; [Van den Berg, Albert; Eijkel, Jan C. T.] Univ Twente, BIOS Lab On A Chip Grp, MESA Inst Nanotechnol, Postbox 217, NL-7500 AE Enschede, Netherlands
资助项目LMB
文献类型期刊论文
条目标识符http://ir.scsio.ac.cn/handle/344004/16230
专题中科院海洋生物资源可持续利用重点实验室
通讯作者shuill@m.scnu.edu.cn
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GB/T 7714
Li, LH,Jin, ML,Sun, CL,et al. High Efficiency Hydrodynamic DNA Fragmentation in a Bubbling System[J]. SCIENTIFIC REPORTS,2017,7:40745-.
APA Li, LH.,Jin, ML.,Sun, CL.,Wang, XX.,Xie, ST.,...&shuill@m.scnu.edu.cn.(2017).High Efficiency Hydrodynamic DNA Fragmentation in a Bubbling System.SCIENTIFIC REPORTS,7,40745-.
MLA Li, LH,et al."High Efficiency Hydrodynamic DNA Fragmentation in a Bubbling System".SCIENTIFIC REPORTS 7(2017):40745-.
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