IMPROVING METHOD OF CHROMOSOMAL DNA EXTRACTION OF BRUCELLA MELITENSIS FOR MULTIPLYING GENE PIECES BY PCR

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PAKZAD I.; REZAEI A.; RASAEI M.J.; ZAVARAN H.A.; KAZEMNEJAD ANOUSHIRAVAN; TABARAEI BAHMAN

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Journal: Daneshvar Medicine: Basic and Clinical Research Journal Year:2004 | Volume:11 | Issue:50 Page(s): 7-12

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Title

IMPROVING METHOD OF CHROMOSOMAL DNA EXTRACTION OF BRUCELLA MELITENSIS FOR MULTIPLYING GENE PIECES BY PCR

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Keywords

BRUCELLA MELITENSISQ4

DNA EXTRACTIONQ2

PCRQ2

L7/L12Q4

Abstract

BRUCELLA MELITENSIS is a gram-negative bacterium, which has a cell wall with a complex structure. Because of cell wall characteristics, the common methods for chromosomal DNA EXTRACTION of gram-negative bacteria have some problems with this bacterium. In this research chromosomal DNA EXTRACTION of this bacterium was carried out for multiplying gene pieces by PCR by CTAB and common method. Because of low efficiency of these two methods, chromosomal extraction of this bacterium was performed by combining this two methods. First, we treated bacteria by SDS and TE, and then we destroyed peptidoglycan by lysozyme. After that, we precipitated protein with phenol-chloroform and polysaccharide by CTAB. In this research, appropriate time for chromosomal extraction from liquid culture was 48 and 72 hours. Then we obtained appropriate amount of 10% SDS and lysozyme respectively 90µ, 85µ (5g/ml). Our results showed that the amount of DNA extracted with this method is higher than the other methodstwo and it reaches 500µ-g/ml. In addition, purity of extracted DNA in this method is better than the other two methods. The extracted chromosomal DNA was used for multiplying L7/L12 (a bacterial ribosomal protein). Because of improved efficiency of this method, it may be used as a method for chromosomal DNA EXTRACTION in clinical samples of blood and milk for diagnosing brucellosis.

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APA: Copy

PAKZAD, I., REZAEI, A., RASAEI, M.J., ZAVARAN, H.A., KAZEMNEJAD, ANOUSHIRAVAN, & TABARAEI, BAHMAN. (2004). IMPROVING METHOD OF CHROMOSOMAL DNA EXTRACTION OF BRUCELLA MELITENSIS FOR MULTIPLYING GENE PIECES BY PCR. DANESHVAR MEDICINE, 11(50), 7-12. SID. https://sid.ir/paper/30428/en

Vancouver: Copy

PAKZAD I., REZAEI A., RASAEI M.J., ZAVARAN H.A., KAZEMNEJAD ANOUSHIRAVAN, TABARAEI BAHMAN. IMPROVING METHOD OF CHROMOSOMAL DNA EXTRACTION OF BRUCELLA MELITENSIS FOR MULTIPLYING GENE PIECES BY PCR. DANESHVAR MEDICINE[Internet]. 2004;11(50):7-12. Available from: https://sid.ir/paper/30428/en

IEEE: Copy

I. PAKZAD, A. REZAEI, M.J. RASAEI, H.A. ZAVARAN, ANOUSHIRAVAN KAZEMNEJAD, and BAHMAN TABARAEI, “IMPROVING METHOD OF CHROMOSOMAL DNA EXTRACTION OF BRUCELLA MELITENSIS FOR MULTIPLYING GENE PIECES BY PCR,” DANESHVAR MEDICINE, vol. 11, no. 50, pp. 7–12, 2004, [Online]. Available: https://sid.ir/paper/30428/en

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