Lecture 1

Question 1

What are pseudogenes

Answer 1

Genes that appear like they are geens for typical functional proteins but in fact code for non fuctional protiens

Question 2

What advantage does polymerase chain reaction have over cloning?

Answer 2

It allows DNA to replicated much faster

Question 3

Other than they typical Polymerase Chain reaction, what are the 2 other types of PCR and what are their advantages?

Answer 3

Reverse Transcriptase PCR which extracts RNA and then converts it to cDNA viathe reverse transcriptase enzyme allowing it to be determined what genes are actually being expressed in a tissue
Real time PCR, this involves the use of a marker allowing you to track the amount of DNA strands made which in turn allows you determine the amount of starting material which means you can gather some information about the gene expression in the tissue

Question 4

What are the 3 steps in PCR

Answer 4

Denaturation, Anealing primers, DNA synthesis

Question 5

What is the most important factor in the denaturation phase?

Answer 5

The temperature to which the solution is raised too must be determined by the enzyme used

Question 6

What is the factor that needs to be regulated closely in the annealing phase?

Answer 6

The lowering of the temperature otherwise, the primers may join to sections of the DNA other than the target sequence resulting in the replication of unwanted material

Question 7

What factor must be tightly contorlled in the DNA synthesis phase of PCR?

Answer 7

The time allowed for elongation as this will control the length of the DNA fragment produced

Question 8

What are the three types of Gel Electophoresis?

Answer 8

Polyacrylamind Gels, Agarose Gel, Pulse-field Gel Electrophoresis

Question 9

What type of Gel electrophoresis uses single stranded DNA of less that 500 nucleotides?

Answer 9

Polyacrylamid Gels

Question 10

What is the range of nucleotide lengths usually used with agarose gel?

Answer 10

300-20,000 nucleotides

Question 11

What type of Gel electrophoresis is used for long DNA strands?

Answer 11

Pulse field gel electrophoresis

Question 12

What is the name of the special nucleotides used for DNA sequencing?

Answer 12

Dideoxynucleotides

Question 13

What 6 molecular techniques rely highly on hybridisation?

Answer 13

Dot Blot, Southern Blot, FISH, SNP arrays, Microarray, Array CGH

Question 14

What is the difference between a dot blot and a southernblot?

Answer 14

In the Dot Blot the hybridisation marker is added directly to the sample to test for its presence while in the southern blot the desired DNA sequence is cut out through use of restriction enzymes

Question 15

What is the purpose of FISH?

Answer 15

To identify potentially harmful mutations that have occured in the cell such as deletions

Question 16

What are SNP arrays used for?

Answer 16

To look for point mutations in order to examine the relationship between this mutation and the observable phenotype

Question 17

What is a microarray?

Answer 17

A microarray is a slate with lots of slots of immobilized known DNA sequences in the form of single stranded molecules, this then allows sample DNA to be washed across the slate so complementary basepair binding will be able to determine if any of the sequences are present

Question 18

What is array CGH used for?

Answer 18

Creation of a karyotype for chromosomal analysis

Question 19

What is the difference between the western and the southern blot?

Answer 19

The southern blot has the ability to test for specific DNA sequences via hybridisation while the western blot uses immunoglobulins to test for specific protiens

Question 20

What is the function of an Immunoassay?

Answer 20

Where antibodies are used to determine the quantity of a specific protein in a cell

Question 21

What is Immunohistochemistry?

Answer 21

A test using immunoglobulins to check for the genexpression of the proteins in the cell