Recombinant DNA Flashcards
1
Q
What is a palindromic DNA sequence
A
A stretch that reads the same (5’ to 3’) on one strand or (3’ to 5’) on the other.
5’-GAATTC-3’
3’-CTTAAG-5’
2
Q
How do restriction enzymes work?
A
They cut dsDNA at palindromic sites.
3
Q
What is hybridization?
A
When two strands of DNA link together due to base pairing between sticky ends.
4
Q
How is hybridization used in gene cloning?
A
A restriction enzyme cuts out a gene of interest at a restriction site, leaves sticky ends.
A plasmid is cut with the same restriction enzyme.
Mix the gene (a DNA fragment) with the plasmid.
Sticky ends allow for hybridization.
DNA ligase seals gene into vector.
Plasmid is inserted into bacteria.
5
Q
What are the steps in PCR?
A
- Denaturation (dsDNA template separates)
- Annealing of primers to ssDNA
- Elongation - polemerase extends primers
Repeat steps 1-3
Results in amplification of template
