AN IMPROVED METHOD FOR RAPID PURIFICATION OF COVALENTLY CLOSED CIRCULAR PLASMID DNA OVER A WIDE SIZE RANGE

被引:3
|
作者
AZAD, AK [1 ]
COOTE, JG [1 ]
PARTON, R [1 ]
机构
[1] UNIV GLASGOW,DEPT MICROBIOL,GLASGOW G12 8QQ,SCOTLAND
关键词
D O I
10.1111/j.1472-765X.1992.tb00698.x
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
An improved method has been developed for the large-scale purification of covalently closed circular (CCC) plasmid DNA molecules of sizes ranging from 4.3 to 73 kb. This protocol uses an alkaline-lysis procedure followed by acid-phenol extraction but with several modifications to previously reported methods. The principal modification is the replacement of NaCl by MgCl2 in the extraction buffer to improve yield and to remove chromosomal and other non-CCC plasmid DNA. Plasmid DNA can be purified in less than 1 h and used successfully in restriction enzyme analysis and cloning experiments.
引用
收藏
页码:250 / 254
页数:5
相关论文
共 47 条
  • [31] Development of a simplified and inexpensive RNA depletion method for plasmid DNA purification using size selection magnetic beads (SSMBs)
    Wang, Xi
    Zhao, Ling
    Wu, Xiaoxing
    Luo, Huaxiu
    Wu, Di
    Zhang, Meng
    Zhang, Jing
    Pakvasa, Mikhail
    Wagstaff, William
    He, Fang
    Mao, Yukun
    Zhang, Yongtao
    Niu, Changchun
    Wu, Meng
    Zhao, Xia
    Wang, Hao
    Huang, Linjuan
    Shi, Deyao
    Liu, Qing
    Ni, Na
    Fu, Kai
    Hynes, Kelly
    Strelzow, Jason
    El Dafrawy, Mostafa
    He, Tong-Chuan
    Qi, Hongbo
    Zeng, Zongyue
    GENES & DISEASES, 2021, 8 (03) : 298 - 306
  • [32] AN IMPROVED METHOD FOR RAPID ISOLATION OF PLASMID DNA FROM WILD-TYPE GRAM-NEGATIVE BACTERIA FOR PLASMID RESTRICTION PROFILE ANALYSIS
    OLSEN, JE
    LETTERS IN APPLIED MICROBIOLOGY, 1990, 10 (05) : 209 - 212
  • [33] Hepatitis B Virus Covalently Closed Circular DNA-Selective Droplet Digital PCR A Sensitive and Noninvasive Method for Hepatocellular Carcinoma Diagnosis?
    Shen, Fan
    Sergi, Consolato
    Sun, Hui-lung
    JOURNAL OF MOLECULAR DIAGNOSTICS, 2018, 20 (03): : 277 - 278
  • [34] RAPID METHOD FOR THE PURIFICATION OF SUPERCOILED PM2 DNA BY AFFINITY CHROMATOGRAPHY ON H1-HISTONE COVALENTLY COUPLED TO AGAROSE
    LAU, PP
    YU, SH
    SPRING, TG
    GRAY, HB
    BIOCHIMICA ET BIOPHYSICA ACTA, 1979, 563 (02) : 313 - 319
  • [35] RAPID METHOD FOR PURIFICATION OF PLASMID DNA AND DNA FRAGMENTS FROM DNA LINKERS USING HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY ON TSK-PW GEL
    HIMMEL, ME
    PERNA, PJ
    MCDONELL, MW
    JOURNAL OF CHROMATOGRAPHY, 1982, 240 (01): : 155 - 163
  • [36] Evaluation of transcriptional efficiency of hepatitis B virus covalently closed circular DNA by reverse transcriptiori-PCR combined with the restriction enzyme digestion method
    Chou, YC
    Jeng, KS
    Chen, ML
    Liu, HH
    Liu, TL
    Chen, YL
    Liu, YC
    Hu, CP
    Chang, CM
    JOURNAL OF VIROLOGY, 2005, 79 (03) : 1813 - 1823
  • [37] Rapid Turnover of Hepatitis B Virus Covalently Closed Circular DNA Indicated by Monitoring Emergence and Reversion of Signature-Mutation in Treated Chronic Hepatitis B Patients
    Huang, Qi
    Zhou, Bin
    Cai, Dawei
    Zong, Yuhua
    Wu, Yaobo
    Liu, Shi
    Mercier, Alexandre
    Guo, Haitao
    Hou, Jinlin
    Colonno, Richard
    Sun, Jian
    HEPATOLOGY, 2021, 73 (01) : 41 - 52
  • [38] A sensitive and accurate quantification method for the detection of hepatitis B virus covalently closed circular DNA by the application of a droplet digital polymerase chain reaction amplification system
    Mu, Di
    Yan, Liang
    Tang, Hui
    Liao, Yong
    BIOTECHNOLOGY LETTERS, 2015, 37 (10) : 2063 - 2073
  • [39] A sensitive and accurate quantification method for the detection of hepatitis B virus covalently closed circular DNA by the application of a droplet digital polymerase chain reaction amplification system
    Di Mu
    Liang Yan
    Hui Tang
    Yong Liao
    Biotechnology Letters, 2015, 37 : 2063 - 2073
  • [40] COVALENTLY CLOSED CIRCULAR DNA OF AVIAN-SARCOMA VIRUS - PURIFICATION FROM NUCLEI OF INFECTED QUAIL TUMOR-CELLS AND MEASUREMENT BY ELECTRON-MICROSCOPY AND GEL-ELECTROPHORESIS
    GUNTAKA, RV
    RICHARDS, OC
    SHANK, PR
    KUNG, HJ
    DAVIDSON, N
    FRITSCH, E
    BISHOP, JM
    VARMUS, HE
    JOURNAL OF MOLECULAR BIOLOGY, 1976, 106 (02) : 337 - 357