MOLECULAR TECHNIQUES

EJC H2 Biology 1 T1 W6 Molecular Biology Techniques Genetics and Inheritance – Molecular Techniques 1. Introduction Molecular biology techniques are common methods used in molecular biology, biochemistry, genetics and biophysics which generally involve manipulation and analysis of DNA, RNA, proteins, and lipids. Since around 1960, molecular biologists have developed methods to identify, isolate, and manipulate mo lecular components in cells including DNA, RNA, and proteins. One important tool used in molecular biology techniques especially in molecular cloning is the restriction enzyme . These enzymes are found in bacteria and provide a defense mechanism against invading viruses. It is an enzyme that cuts DNA at or near specific recognition nucleotide sequences known as restriction sites. Several techniques used in the field of molecular biology include:  Polymerase chain reaction (PCR)  Gel electrophoresis  Macromolecule blotting and probing e.g. Southern blotting , Northern blotting, Western blotting and Eastern blotting  Expression cloning 2. Learning Outcomes (k) describe the principles and procedures of these molecular techniques: i. polymerase chain reaction (including its advantages and limitations) ii. gel electrophoresis iii. Southern blotting and nucleic acid hybridisation Use the knowledge gained in this section in new situations or to solve related problems. 3. References i. T.A.Brown (2010) Gene cloning & DNA Analysis, 6 th Edition, Wiley -Blackwell. Chapter 1 to 7, 9. ii. Watson, Mayers, Caudy, Witkowski (2007), Recombinant DNA, Genes and Genomes – A short Course. Cold Spring, Harbor Lab Press. Chapter 4 iii. Campbell and Reece (2011) Biology, 9th Edition, Pearson Education. Chapter 20. iv. Turner, McLennan, Bates and White (2001) Molecular biology, 2 nd Edition, BIOS. Sections G, H, I and J. v. Green, Stout and Taylor (1997), Biological Science Volumes 1 and 2, 3rd Edition. vi. Hoh Yin Kiong (2003), Longman A-Level Course in Biology, Core Syllabus Volume 1 (for AS and A-Level) vii. Weaver (2008), Molecular Biology(4th ed.). McGraw-Hill. viii. http://www.dnalc.org/resources/spotlight/index.html (Cold Spring Harbor Laboratory DNA Learning Centre. Look for Polymerase Chain Reaction).

EJC H2 Biology 2 T1 W6 Molecular Biology Techniques Contents 1. Introduction ............................................................................................................... 1 2. Learning Outcomes.................................................................................................... 1 3. References ................................................................................................................. 1 4. Molecular Biology Techniques .................................................................................. 3 A. Polymerase Chain Reaction………………………………………………….…………………………...3 B. Gel Electrophoresis………………………………………………………………………………..12 C. Southern Blott