Molecular Diagnosis Flashcards
How is our own DNA protected from restriction enzymes?
Methylation.
How can we isolate particular pieces of DNA from a sequence?
Use multiple restriction enzymes.
What is DNA electrophoresis?
It is a method which can be used for visualising colourless DNA. Fragments are separated using a negative charge. Fragments have to move on the plate so larger molecules move less far, gradient of molecule sizes created.
What are the four requirements for gel electrophoresis?
Gel - allows separation, Buffer - maintains charge, Power supply - creates charge difference, stain - visualise DNA.
What is the function of DNA ligase in molecular diagnosis?
When complementary base sequences align, this can lead to the formation of new bonds.
What is used in gene cloning?
Plasmids from Bacteria are used in gene cloning.
What is recombinant DNA?
This is the plasmid ring with the DNA in it as well.
What is the use of cloning genes?
We can use cloned genes in order to make useful proteins such as insulin.
What is genetic screening?
This is where we are looking for a specific gene. To look at this properly we clone genes.
What is PCR?
Polymerise chain reaction. It is a method for obtaining large amounts of a particular piece of DNA for analysis.
What is a primer?
This is a short molecule which is complementary to part of the DNA sequence. It is a site where DNA replication begins.
Explain the cycle process of PCR.
First DNA is heated to 95 degree so that it is denatured. It is then cooled to 60 so that primers anneal and then heated to 75 when replication takes place. The DNA is then reheated.
What is used so that DNA polymerase denaturing is not a problem in PCR?
Taq, thermostable DNA polymerase so that it does not have to be replaced with each cycle.
What is the use of PCR?
PCR allows amplification of a specific piece of DNA which can then be investigated for base mutations or deletions/insertions.
State the difference between protein and DNA gel electrophoresis.
Protein gel electrophoresis is done vertically rather than horizontally so proteins move down the plate.