Dna Analysis Practical Write-Up
Title: DNA analysis
Aim: a) Isolate and Purify Bacterial Chromosomal DNA from a strain of E.coli
b) Visualization of restriction fragments by Agarose Gel electrophoresis
Objectives: * to isolate and purify bacterial chromosomal DNA from a strain of E.coli * to analyze and identify DNA by use of a spectro-photometer * to use restriction enzymes to cleave DNA into fragments * to visualize the restriction fragments by gel electrophoresis * to compare the different DNA fragments generated by use of molecular markers
Abstract
This work describes a lysis method for the isolation and purification of bacterial genomic DNA and visualization of the restriction fragments by agarose gel electrophoresis. It was noted that for one to isolate and purify bacterial chromosomal DNA several steps are taken into consideration. DNA was found to absorb at 260nm wavelength in a UV spectrophotometer. Restriction enzymes were added to cleave DNA which would produce various DNA fragments. DNA can be separated into different sized fragments by gel electrophoresis. The bacterial DNA was successfully isolated and purified however it could not be observed after running the gel.
DNA analysis is a standard practice for defining paternity or maternity, predisposition to disease, embryonic health and criminal guilty. But in our context, DNA analysis is mainly used for predisposition of diseases in bacteria. Bacteria are pathogenic microorganisms that cause infectious diseases including cholera, syphilis, anthrax and leprosy. The most common fatal bacterial diseases are respiratory infections such as tuberculosis (Barnum S.R; 1998).
Nucleic acids encode information relating to cell structure and function. Cells have the ability to make copies of their DNA and pass this information to daughter cells. Nucleic acids are polymers of nucleotides. Nucleotides are composed of ribose (a 5` carbon) sugar and either a purine and pyrimidine base at 1` position. The
Aim: a) Isolate and Purify Bacterial Chromosomal DNA from a strain of E.coli
b) Visualization of restriction fragments by Agarose Gel electrophoresis
Objectives: * to isolate and purify bacterial chromosomal DNA from a strain of E.coli * to analyze and identify DNA by use of a spectro-photometer * to use restriction enzymes to cleave DNA into fragments * to visualize the restriction fragments by gel electrophoresis * to compare the different DNA fragments generated by use of molecular markers
Abstract
This work describes a lysis method for the isolation and purification of bacterial genomic DNA and visualization of the restriction fragments by agarose gel electrophoresis. It was noted that for one to isolate and purify bacterial chromosomal DNA several steps are taken into consideration. DNA was found to absorb at 260nm wavelength in a UV spectrophotometer. Restriction enzymes were added to cleave DNA which would produce various DNA fragments. DNA can be separated into different sized fragments by gel electrophoresis. The bacterial DNA was successfully isolated and purified however it could not be observed after running the gel.
DNA analysis is a standard practice for defining paternity or maternity, predisposition to disease, embryonic health and criminal guilty. But in our context, DNA analysis is mainly used for predisposition of diseases in bacteria. Bacteria are pathogenic microorganisms that cause infectious diseases including cholera, syphilis, anthrax and leprosy. The most common fatal bacterial diseases are respiratory infections such as tuberculosis (Barnum S.R; 1998).
Nucleic acids encode information relating to cell structure and function. Cells have the ability to make copies of their DNA and pass this information to daughter cells. Nucleic acids are polymers of nucleotides. Nucleotides are composed of ribose (a 5` carbon) sugar and either a purine and pyrimidine base at 1` position. The
References: 1. Allison L.A. (2012). Fundamental Molecular Biology, 2nd edition. Denvers. John Wiley and Sons Inc. 2 3. Besty Tom and Keogh Jim, (2005), Microbiology demystified, New York; MacGraw-Hill. 4. Cowan Majorie Kelly, (2009), Microbiology: A Systems Approach, 3rd edition; New York; MacGraw-Hill. 5. Gupta, P.K. (2006). Elements of Biotechnology, Meerut. Rastogi Publications. 6. Stryer L, Berg J.M and John Tymozcko. (2006). Biochemistry. 5th edition. California. W.H Freeman and Company. 7. Wiser, M.F. (2002). Methods in cell biology. Berlin. Springer Verlog