UNIT 4. Techniques used in Molecular Biology. Objectives. On completion of this unit students will be able to: Outline the steps in Polymerase Chain Reaction Analyse DNA agarose gel electrophoretograms Describe DNA hybridization and its application in probe synthesis
Techniques used in Molecular Biology
On completion of this unit students will be able to:
The cycling reactions :
54°C :hydrogen bonds are constantly formed and broken between the single stranded primer and the single stranded template.
The use of a thermostable polymerase allows:
Is there a gene copied during PCR and is it the
Factors that affect yield:
The product is of the right size:
Factors that affect specificity:
1. The method is especially useful for searching out disease organisms that are difficult or impossible to culture:
2. PCR can also be more accurate than standard tests.
3. The method is also leading to new kinds of genetic testing.
These tests diagnose not only people with inherited disorders, but also people who carry deleterious mutations that could be passed to their children.
4. PCR can provide enormous peace of mind to people who are trying to have children-
The rate of migration through the electric field
Steps in Southern blotting:
The strand synthesis reaction: