8-21 DNA technology Flashcards
What is a blunt end?
When DNA is cut straight through the double strand without seperating bases
What is the process of transformation?
First increase the permeability of the membranes, e.g. use calcium ions for bacteria
Keep cold to reduce metabolic activity
Once its exposed quickly raise the temperature to 42 (heat shock)
This causes the plasmid to be quickly expadited into the bacteria
This encourages the movement of the vector into the host cell
What are the problems that can occur during insertion?
The DNA fragments can join up with each other because it is looking for complimentary pairs
Sometimes plasmids close up before the gene has been inserted because the sticky ends are complementary
Sometimes two plasmids join
What are the advantages of recombinant DNA technology?
GMO plants produce plant organs which can then be harvested
Replacing defective genes can be used to cure genetic disorders
GMO crops can survive harsher conditions
What are the risks of recombinant DNA technology?
Can a company patent a gene and own DNA?
Could a new disease be created accidentally?
People could change their genetic makeup (eugenetics)
What are DNA probes?
Short single strands of DNA which contain a DNA marker
What are radioactively labelled probes?
Made up of nucleotides with the isotope 32P
Identified using an x-ray film
What are fluorescently labelled probes?
Probes which emit light
What is the process of DNA hybridisation?
Seperate the two strands of DNA
Add complimentary DNA from someone else to see if they’re complimentary
What is hybrid DNA?
A form of recombinant DNA where the complimentary parts of the strand anneal
What is recombinant DNA?
DNA from two different source
What does the annealing of probes do?
It confirms the base sequences
What can DNA be split up by?
Heat or DNA helicase
What is the function of PCR?
It amplifies your sample of DNA
How long is the DNA sample in PCR?
An entire genome or a fragment
What does PCR stand for?
Polymerase chain reaction
What is required for PCR in terms of DNA polymerase?
DNA polymerase will denature at higher temperatures
What is required for PCR? (without explanation)
DNA polymerase
pH buffer
Thermal cycle
Primers
Free nucleotides
What is required for PCR in terms of the thermal cycler?
All ingredients are put into a thermal cycler which changes the temperature
What are primers?
Short sequences of nucleotides that have a set of bases that are complementary to the DNA fragments