Biotechnology Flashcards
What is biotechnology?
Any technique applied to biological systems or living organisms to make
or modify products or processes for a specific purpose
What is DNA technology?
Techniques used to isolate, purify, analyze, and manipulate DNA to serve a
specific purpose
What is genetic engineering?
Altering genes of genomes for practical purposes
What are the steps in DNA cloning?
- Identify a recognition
- Add restriction endonuclease
- Sticky ends result
- Insert genes into the plasmid
- Transformation and cloning
What two main components are used in creating recombinant DNA?
Bacterial plasmid DNA (vector dna), gene of interest
What are the steps in creating recombinant DNA?
- put the gene we want to know about into the plasmid
- put the plasmid into the bacterium
- the gene can be transcribed/translated and replicated when the bacterium divides
What type of bacterium should they use for recombinant DNA?
plasmid-free bacteria
What sites were bacterial plasmid DNA used in labs engineered to have?
multiple cloning site
What are the steps in polymerase chain reaction? (PCR)
- denaturing
- primer annealing
- extension
- repeat steps 1-3
What components of normal DNA replication are missing in PCR?
helicase, ligase, primase, single strand binding protein, etc
What components are in PCR?
DNA, taq polymerase, primers, dNTPs
Why is Taq polymerase used in PCR?
can withstand the high temperatures
What jobs does the denature stage of PCR do?
helicase, topoisomerase, SSBP
Are RNA primers used in PCR? Why?
no, because they can’t be removed easily so we use DNA primers so they don’t have to be
What does the length of the bands in electrophoresis represent?
longer band means more base pairs, shorter means less
Which fragment will move further in electrophoresis, one that is 200bp and one than is 500bp?
200bp because its shorter
What is a DNA marker in electrophoresis?
the fragments in the marker are a known length so we can use to tell the approx size of samples
What does a brighter band mean in electrophoresis?
more dna
Once we select the plasmid and amplify our gene of interest how can we insert this gene into the plasmid?
cut plasmid DNA with enzymes
What is a palindromic sequence? example?
sequence that is reversible
racecar
Where are restriction sites located?
multiple cloning sites
What are sticky bonds?
short, single stranded ends of dna after being cut by restriction endonuclease that can form hydrogen bonds with complementary sticky ends
Do the plasmid and gene of interest need to be cut with the same enzyme?
yes
What is used to connect sticky ends? What is this process called?
DNA ligase
ligation
