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International Journal of Pure & Applied Bioscience (IJPAB)
Year : 2018, Volume : 6, Issue : 5
First page : (533) Last page : (541)
Article doi: : http://dx.doi.org/10.18782/2320-6988

A Comparative Study on Different Plasmid Isolation Procedures

Utsa Roy^1#*, Archana Mishra^1#, Prasanta Jana² and Sutanu Karmakar³

¹Fish Genetics and Biotechnology Division, ICAR-CIFE, Mumbai
²Fish Nutrition, Biochemistry and Physiology Division, ICAR-CIFE, Mumbai
³Aquatic Environment and Health Management Division, ICAR-CIFE, Mumbai
#- Authors contributed equally in this study
*Corresponding Author E-mail: utsaroy.007@gmail.com
Received: 18.07.2018 | Revised: 24.08.2018 | Accepted: 30.08.2018  

 ABSTRACT

Plasmid DNA has several applications in the field of modern biotechnology. Despite the presence of several methods to remove contaminants and/ or to purify plasmid DNA from primary isolate, the first technique of plasmid isolation determines the quality of the final product. In this study, some popular methods of plasmid DNA isolation have been compared in terms of their rapidity, plasmid quality and ease of experiments. Three different STET buffers used in boiling lysis process were also compared to identify the best performing one. Highest quantity of RNA contaminants, as well as plasmid, was isolated in the alkaline extraction process. Best supercoiled plasmid percentage was isolated by boiling lysis STET buffer 2. The RE digestion was non-reactive/ improper in boiling lysis methods whereas it was perfect in alkaline lysis despite huge quantity of RNA contaminants. Alkaline lysis was proved to be the best one among all and with some further purification processes indicated, other processes can also be used efficiently.

Key words: Alkaline lysis, Boiling lysis, STET buffer, Plasmid isolation, Plasmid quality

Abbreviations-
BL1: Plasmid isolation by boiling lysis process with STET 1 buffer
BL2: Plasmid isolation by boiling lysis process with STET 2 buffer
BL3: Plasmid isolation by boiling lysis process with STET 3 buffer

Full Text : PDF; Journal doi : http://dx.doi.org/10.18782

Cite this article: Roy, U., Mishra, A., Jana, P. and Karmakar, S., A Comparative Study on Different Plasmid Isolation Procedures, Int. J. Pure App. Biosci.6(5): 533-541 (2018). doi: http://dx.doi.org/10.18782/2320-7051.6988