Restriction Enzymes & Cloning Flashcards
1
Q
Define Recombinant DNA technology
A
the ability to manipulate DNA with precision in a test tube or an organism
2
Q
Define Cleavage of DNA
A
Carried out at specific sites by restriction nucleases,
3
Q
Define DNA Ligation
A
allows for seamless joining of DNA molecules
4
Q
Define DNA Cloning
A
- done through cloning vectors or polymerase chain
- a portion of the genome is ‘purified’ away from the remainder of the genome by repeatedly copying git to generate many billions of identical molecules
5
Q
Describe Restriction Enzymes
A
- purified from bacteria
- cuts the DNA double helix at specific sites defined by local nucleotide sequence
6
Q
What are the functions of restriction enzymes ?
A
to cleave DNA at sequence specific sites, producing DNA fragments
7
Q
Describe DNA molecular Cloning
A
- any DNA fragment can be cloned
- two senses of the term
1. act of making many identical copies of a DNA molecule
2. the isolation of a particular stretch of DNA from the rest of the genome
8
Q
What are the 7 steps of Molecular Cloning ?
A
- choice of host & cloning vector
- prep of vector DNA
- Prep of DNA to be cloned
- creation of recombinant DNA
- intro of recombinant DNA into host
- selection of organisms containing recombinant DNA
- Screening of clones with desired DNA & traits
9
Q
How do you isolate a gene of interest ?
A
- cells are lysed and plasmid DNA is isolated
- cloned DNA fragments can be readily recovered via cutting it out using plasmid DNA
- separation of plasmid DNA by gel electrophoreses
