Molecular Diagnostics

What are two of the main techniques that can be used in molecular diagnostics?

What are the three simple steps which describe how diseases can be diagnosed by analysis of DNA from a patient?

What is the function of restriction enzymes?

What are the two types of ends that can result from cutting with restriction enzymes?

What does it mean when it says restriction enzyme cutting sites are pallindromic?

Give 2 examples of restriction enzymes.

Does Eco RI cleave DNA to sticky ends or blunt ends?

What is the size of the target site recognised by Eco RI?

What is the sequence that Eco RI recognises?

How large would the fragments produced by Eco RI be if it cut human DNA? How many fragments would be produced if it digested human DNA?

What is the function of gel electrophoresis?

Which end do DNA and RNA migrate to in gel electrophoresis?

Which migrate faster in gel electrophoresis: large or small fragments?

Does the restriction enzyme Hae III produce blunt ends or sticky ends?

How can DNA/RNA fragments be seen as bands on the gel?

What is used in southern blotting to show the fragment of DNA that is needed following electrophoresis?

What are the main steps in southern blotting?

How does the DNA from the gel blot onto the nylon membrane?

What is it important to do after the probe and nylon membrane are incubated overnight?

What is the restriction enzyme used in diagnosis of sickle cell via southern blotting?

What is it callled when digestion of normal beta-globin genes and mutated ones (in sickle cell) with Mst II produces fragments of different sizes?

Is northern blotting the same or different to southern blotting?

What is the function of PCR?

What are three circumstances where PCR may be used?

What are two of the key components of PCR?

True or false: two primers are needed in PCR - a forward and a reverse.

What are the four things that a PCR reaction contains?

In most cases how many cycles of amplification occur? How long may this take?

What are the three main steps in PCR?

What are the three main steps in PCR?

True or false: the amount of target fragment triples during each cycle there after n cycles there will be 3^n copies of the gene.

What kind of mutation occurs in huntingtons disease? Give examples.

When can PCR be used to show that someone has the huntingtons gene?

What are the three main steps involved when PCR is used?

What form of PCR is often used to diagnose/find bacteria and viruses? Why and how?