Biotechnology Flashcards
Define biotechnology
Use of microbiological and biochemical techniques to produce useful products and solve practical problems
Name some examples of biotechnology
Identification technologies and DNA cloning
Describe DNA sequencing
Determining the sequence of bases in a whole genome
Describe DNA probes
Short ,labeled single strand of bases used to detect a complementary sequence of DNA
You look for the fluorescence in the sample using the fluorescent microscope to view if it’s positive or negative
Is colony blotting used in identification technologies
Yes
What is FISH
Fluorescence in situ hybridization
Describe the process of FISH
Treat the cells with a chemical. Opens membranes to allow probes to enter. Probe sneaks in to entact cells. Put florescence finds the DNA binds and then you can look for it with the microscope
What can DNA microarrays do
Detect many genes at one time
Describe the process of microarrays
You start with the chip add holes with the DNA probe looking for something. The chip is already embedded within the spots. Have to add DNA that has fluorescent label on it. Wash the unknown with the probe. One sample goes through the probe you put it through the computer to read the colors
Describe DNA fingerprinting
Isolation and visualization of DNA sequences to help identify an unknown
Describe restriction fragment length polymorphisms (RFLP)
It’s the creation of fragments of different lengths using restriction enzymes
What do restriction enzymes do
Cut DNA at specific 4-6 base pair sequences
Describe gel electrophoresis
Visualize fragments using gel electrophoresis. The DNA gel is stained with ethidium bromide, a dye that fluoresces under the UV light
If it’s a short fragment it’s going to go fast and long fragments is the opposite that’s how they’re separated
What is the size standard used for
A control (molecular weight ladder)
Describe the polymerase chain reaction(PCR)
The amplification of a DNA sequence using specific primers and the thermal cycler to replicate DNA.
It’s used to visualize amplified DNA on a gel.
The band is thick because they’re all the same size
What is DNA cloning
The procedure in which a gene of interest is extracted and inserted into another cell where he can be expressed and replicated
What does DNA cloning involve
Recombinant DNA(rDNA), genetic engineering
What is rDNA
DNA formed from joining DNA from two sources
What is genetic engineering
When you alter an organism by changing its DNA
Name the steps for DNA cloning
1) obtain DNA
2) Cut DNA at restriction sites
3) insert gene of interest into vector
4) insert vector into hosting cell(mass Producer)
5) collect product
What happens in step one of DNA cloning
Lyse source cell with gene of interest and collect DNA
Describe step two of DNA cloning
Restriction enzymes create multiple fragments of variable length with sticky ends
Describe step three of DNA cloning
Vectors: plasmids, phages
Cut vector open with same restriction enzyme
Describe step four of DNA cloning
Hosts: E. coli, bacillus subtilius
Methods: artificial transformation(plasmids) - heat shock and electroporation Artificial transduction(phages)
Culture bacteria
Describe step five of DNA cloning
Protein
- lyse cells and purify protein
- collect and purify secreted proteins from cells
Gene
-lyse cells and collect gene of interest
What are variations of DNA cloning
Eukaryotic genes into prokaryotic hosts
-create cDNA from mRNA
Describe the ways that genes are put into eukaryotic hosts
Host cells grown in culture
DNA insertion methods
Name the DNA insertion methods
Artificial transformation
Viral or bacterial-mediated gene delivery
Gene gun (plant cells)
Microinjection (animal cells)
What are transgenic organisms
Organism has foreign DNA present in every cell
Also called genetically modified organisms(gMO’s)
What are four ways that animal viruses can interact with host cells
Acute ,chronic ,transformation, Latent reverse transcription and special Enzyme
