Advances in Molecular Biology Tools: Genetic Engineering Techniques and Applications
This lecture discusses the evolution of genetic engineering techniques from traditional breeding methods to advanced molecular biology tools. It highlights how technology, such as automated cell sorting and restriction enzymes from bacteria, has revolutionized DNA analysis and manipulation. Key methods like gel electrophoresis, cloning, sequencing, and the use of DNA microarrays are explored to demonstrate their importance in modern genetic research. Understanding these tools is essential for advancements in genetics and biotechnology.
Advances in Molecular Biology Tools: Genetic Engineering Techniques and Applications
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Presentation Transcript
Genetic Engineering 1 Lecture 18 Pages 323 - 340
Old fashioned way was to breed for what you wanted 10_01_experiment.DNA .jpg Man has been doing this since the Agricultural revolution, some 10,000 years ago… The Chinese have been doing it! Mendel did it too!
What do we do NOW… • Breeding is still used. • However, quicker and more results orientated methods now exist due to technology. • First, we may need to filter the cell types… • Label target cells with florescent dye attached to characteristic cell specific proteins • Brain cells have some unique proteins • Liver cells have other unique proteins, etc.
Still used - an automated cell sorter 10_02_cell_sorter.jpg
Once you have the cells what next? • OPTIONS • Issue was to examine the DNA in a consistent manner • Best method (Restriction Digestion) is to use restriction enzymes (REs) • Come mainly from bacteria • They are bacterial immune systems against foreign DNA • They are all Proteins • Many types have been isolated from different species of bacteria • Scientists use REs individually or in a mix
What do you do with these digested fragments of DNA? • Isolate those that you want to work on • How? • Best method is to use gel electrophoresis • Agarose • Polyacrylamide
Then what do you do with this piece of DNA? • EITHER • Clone • Sequence • Rely on the use of dideoxy nucleotides
Other methods used Shotgun cloning In the Dark…
10_09_Shotgun.sequenc.jpg Use computer software to assemble
Problems with this method What can they be?
YACsBACs Cloning Techniques used to obtain DNA sequence information
Blotting Purpose = to make a permanent record of the results of a gel electrophoresis. The compass - • Southern blots - DNA • Northern blots - RNA • Western blots - Proteins • Eastern blots - ?
DNA microassays New techniques Many uses DNA Chips
