Biotechnology Flashcards
1
Q
What is recombinant DNA?
A
a DNA strand that is created using DNA pieces from 2+ sources
2
Q
What is a restriction enzyme? How are they recognized?
A
an enzyme that cuts DNA at a specific location in a base sequence; palindromes
3
Q
What is a recognition site?
A
a sequence of bases on a DNA strand that restriction enzymes bind to
4
Q
What is the point of bacterial transformation?
A
Express gene in other organizations
5
Q
What is the point of PCR?
A
Amplify small amounts of DNA
6
Q
What is the point of gel electrophoresis?
A
Separate DNA after recombining to see results
7
Q
What is the point of recombinant DNA?
A
Combine DNA for a gene
8
Q
What happens during bacterial transformation?
A
- DNA is put into a plasmid to transfer genes
- CaCl2 is to neutralize charge of membrane and DNA
- Ice and heat shock (42 C) causes convection currents to force plasmid into cell
- LB and 37 C is optimal conditions for growth for bacteria to survive ampicilin
9
Q
What happens during PCR?
A
- Heat to 95C to cause H bonds to break between complementary bases
- Cool to 50-65 C allows DNA primers to anneal with complementary base pairing to target area
- Heat to 72C allows Taq polymerase to function optimally to elongate primers adding nucleotides
- Repeat until multiple copies of target area is made
10
Q
What happens during gel electrophoresis?
A
- Gel hardens and is placed in electrolyte solution (DNA is negatively charged)
- Comb is placed in liquid to produce sample wells
- DNA is placed in wells and moves to positive pole as soon as current is run through electrolyte solution
- The smaller the molecule, the faster it runs
- Molecular weight marker is fragment of known sizes to be used as comparison
11
Q
What happens during recombinant DNA?
A
- Restriction enzymes cut plasmid
- You add the fragment of DNA with the gene you wish to insert; you may use PCR to amplify the amount
- Use DNA ligase to combine
- Use gel electrophoresis to see which fragments you got
