Biochemistry 2280A Chapter 10: Topic 21 - RECOMBINANT DNA TECHNOLOGY TOOLS Learning Objectives

• School: Western University
• Department: Biochemistry
• Course: Biochemistry 2280A
• Professor: Susan Meakin

Document Summary

Describe the tools of recombinant dna technology and explain how each functions. Cut double stranded dna at specific sequences (usually palindromic symmetrical around a central point) Sticky ends short ss overhangs help the cut dna molecules join back through complementary base-pairing. Ecorl (gaattc) and hindlll (aagctt: gel electrophoresis. Method used to separate dna fragments on the basis of their size. Mixture of dna fragments loaded at one end of a slab of agarose/polyacrylamide gel which contains network of pores. Voltage is applied; -ve dna gragments migrate towards +ve electrode. Larger fragments migrate more slowly since their progress is impreded more by the gel matrix: nucleic acid hybridization. Aka dna renaturation is driven by re-formation of the h bonds between complementary bp. Ss dna probe that"s complementary to the nucleotide sequence of interest is designed carry a fluorescent/radioactive label: northern blotting/southern blotting. Mix of ds dna fragments separated according to length by gel electrophoresis.