Structural Biochemistry/DNA manipulation techniques
Different techniques are commonly used to determine or alter DNA sequences. The most important ones include:
- DNA sequencing: It provides us a lot of information about gene architecture, the control of gene expression, and protein structure.
- solid state synthesis of nucleic acids: The desired sequences of nucleic acids can be synthesized precisely de novo and used to identify or amplify other nucleic acids.
- restriction enzyme analysis: These Restriction Endonucleases provides an investigator to manipulate DNA segments by cutting them from the recognized sites.
- polymerase chain reaction (PCR) : This is one of the most useful methods to amplify DNA sequence billion fold. This technique requires denaturing of DNA and hybridizing primers complementary to strand forward and reverse and the DNA is synthesized on both strands in the 5' to 3' direction. This technique is used to recognize pathogens and genetic diseases, to determine the source of a hair left at the scene of a crime, and to resurrect genes from fossils.
- Blotting techniques: Southern and Northern Blots are used to separate DNA and RNA, respectively.1
1. Berg, Jeremy M. 2007. Biochemistry. Sixth Ed. New York: W.H. Freeman. 68-69, 78. 2. Voet, Voet, Pratt (2004). - Fundamentals of Biochemistry