DNA Manipulation Flashcards
What is gene technology?
It is a broad field which includes analysis of DNA as well as genetic engineering and other forms of genetic modification
What does GMO stand for?
Genetically modified organisms
What does genetic engineering refer to?
Refers to the artificial manipulation of genes: adding or subtracting genes or changing the ways genes work
How can gene technologies benefit humanity?
- increasing crop population
- increasing livestock production
- preventing/ fighting disease
- reducing pollution and waste
What is recombinant DNA?
a piece of DNA that has been altered by adding a new section of DNA usually from a different organism
What do restriction enzymes allow?
Allows genetic engineers to cut up DNA in a controlled way
Where does a restriction enzyme cut?
At the specific recognition site
An example of recombinant DNA molecules include molecules formed by?
the splicing of DNA from one species into that of a second species.
Another name for a restriction enzyme is?
Endonuclease
Each restriction enzyme cuts DNA at particular sequence of bases called the?
Recognition sequence
Amplification of DNA in the polymerase chain reaction requires?
DNA polymerase
What is the name of the enzyme that is used to join fragments of DNA from different sources?
ligase
What is annealing?
When two matching ‘sticky ends’ come together they join by base pairing
What is a plasmid?
Circles of DNA found within bacterial cells
Is the bacterial plasmid seperate from the DNA responsible for the bacterias phenotype?
Yes
What are plasmids used for?
To produce specific proteins for the bacteria
What is electrophoresis?
A technique that seperates large molecules (nucleic acids or protein) on the basis of their size, electric charge and physical properties
What is the gel covered in, in gel electrophoresis?
A buffer solution
What is the purpose of a buffer solution?
- stop gel from melting
- allow current to move along the gel
- maintain pH of DNA
Do large molecules move faster or slower through the gel?
Slower
What gel is used in electrophoresis?
Agrose gel
What does PCR stand for?
Polymerase chain reaction
What is pcr?
A process of DNA replication in a laboratory setting
What does the process of pcr require?
- free nucleotides
- original DNA sample
- rna/ DNA primers
- taq polymerase
Where does taq polymerase come from?
A bacterial enzyme from hot springs
What is a vector?
A self replicating DNA molecule used to transmit a gene from one organism into another
What properties must vectors have?
- be able to replicate in host
- have on or more sites at which a restriction enzyme can cut
- have a genetic marker so it can be identified
Are plasmids stable?
Yes
Why are plasmids stable?
Cause they are circular and can easily be transferred form one bacteria to another
What is a plasmid that has been genetically altered called?
Recombinant DNA
What is the organism called when recombinant DNA which was a genetically altered plasmid is paced in a bacteria?
Transgenic organism
What is a transgenic organism?
Are species that have been modified by bringing in the DNA of a different species
What is the difference between a GMO and transgenic organism?
Transgenic organisms bring in DNA from another species whereas gmos are organisms that have had their own DNA altered
What are ways a plasmid can be transferred to a bacteria?
- transformation solution
- shock treatment
- agar pates
What is a transformation solution?
A solution usually calcium chloride that the bacteria is placed in
What does the transformation solution do?
Makes the bacteria’s cellular structure more permeable and plasmids can move in and out more easily
What is the process of shock treatment?
The bacteria and plasmids are mixed together in test tubes, then placed in ice baths, then warmed and back into ice
What is the purpose of shock treatment?
Allows bacteria to take up plasmids
What is the process using agar plates?
Bacteria are placed on different agar plates these plates are treated so bacteria growth can be analysed
What is rational drug design?
The process of designing drugs that will have an effect on a normal cellular process
What is a well in electrophoresis?
Holes made in the gel with a comb acting as a reservoir for the DNA solution
What is a DNA marker in gel electrophoresis?
A mixture or DNA molecules with the known molecular weights (size) are often run in one lane
What is the purpose of DNA markers in gel electrophoresis?
They are used to estimate the size of the DNA in the sample lane
What charge does DNA have?
Negatively charges because of the phosphate
What is the first step of pcr?
The DNA sample is heated to temps around 98 so they are denatured and strands seperate
What is the second step of pcr?
The sample is cooled and primers anneal to strands
What is the third step of pcr?
Free nucleotides and the enzyme taq polymerase are added at 78 degrees. The taq polymerase binds to the primers and extends
After one cycle how many strands of DNA are there?
2 copies of original DNA
During an experiment a controlled variable is kept?
Constant
When graphing results what axis is the independent variable found on?
Horizontal
When graphing results what axis is the dependent variable found on?
Vertical
During PCR what temperature is ideal for the extension stage?
72 degrees
What is the purpose of a primer?
To act as a short sequence of nucleotides that provides a starting point for DNA synthesis
What is a non ethical consideration of genetic screening of newborn babies?
The cost
