Fergus - Recombinant DNA Flashcards
What is recombinant DNA technology?
(3 Definitions)
The ability to clone and manipulate DNA sequences
The ability to join 2 pieces of DNA from different organisms
The ability to generate proteins from recombinant DNA sequences in organisms that would not normally produce those proteins
What is recombinant DNA?
DNA that has been formed artificially by combining constituents from different organisms
How is recombinant DNA made?
(5)
Two pieces of DNA
An endonuclease (BamHI) is used to digest/break up the DNA
This creates two sticky ends which then join together
This is then sealed with DNA ligase
This creates recombinant DNA
How many types of restriction endonucleases are there?
Three types
Write a note on type 1 restriction endonucleases.
(3)
Multi subunit
Cleave at random up to 1000 bps from recognition sequence
Methylase and endonuclease activity
Write a note on type 2 restriction endonucleases
(3)
Single subunit (endo and methyl activity in the one unit)
Cleave DNA within recognition sequence
Most common
Write a note on type 3 restriction endonucleases
(3)
Multi subunit
Endonuclease and methylase activity
25 bps from recognition sequence
What is Hae III?
(3)
A restriction enzyme that searches the DNA molecule until it finds a specific sequence of four nitrogen bases
5’ GGCC 3’
3’ CCGG 5’
It creates blunt ends
What is a sticky end?
A staggered cut
At what temperature is DNA denatured?
95 degrees
At what temperature are primers annealed?
Between 50 and 70 -> depending on temperature
At what temperature does TAQ work at?
72
