Lecture 1 Flashcards
What are pseudogenes
Genes that appear like they are geens for typical functional proteins but in fact code for non fuctional protiens
What advantage does polymerase chain reaction have over cloning?
It allows DNA to replicated much faster
Other than they typical Polymerase Chain reaction, what are the 2 other types of PCR and what are their advantages?
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
What are the 3 steps in PCR
Denaturation, Anealing primers, DNA synthesis
What is the most important factor in the denaturation phase?
The temperature to which the solution is raised too must be determined by the enzyme used
What is the factor that needs to be regulated closely in the annealing phase?
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
What factor must be tightly contorlled in the DNA synthesis phase of PCR?
The time allowed for elongation as this will control the length of the DNA fragment produced
What are the three types of Gel Electophoresis?
Polyacrylamind Gels, Agarose Gel, Pulse-field Gel Electrophoresis
What type of Gel electrophoresis uses single stranded DNA of less that 500 nucleotides?
Polyacrylamid Gels
What is the range of nucleotide lengths usually used with agarose gel?
300-20,000 nucleotides
What type of Gel electrophoresis is used for long DNA strands?
Pulse field gel electrophoresis
What is the name of the special nucleotides used for DNA sequencing?
Dideoxynucleotides
What 6 molecular techniques rely highly on hybridisation?
Dot Blot, Southern Blot, FISH, SNP arrays, Microarray, Array CGH
What is the difference between a dot blot and a southernblot?
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
What is the purpose of FISH?
To identify potentially harmful mutations that have occured in the cell such as deletions
What are SNP arrays used for?
To look for point mutations in order to examine the relationship between this mutation and the observable phenotype
What is a microarray?
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
What is array CGH used for?
Creation of a karyotype for chromosomal analysis
What is the difference between the western and the southern blot?
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
What is the function of an Immunoassay?
Where antibodies are used to determine the quantity of a specific protein in a cell
What is Immunohistochemistry?
A test using immunoglobulins to check for the genexpression of the proteins in the cell
