Transcript Biotechnology: Tools and Techniques of the Trade

What can you do with DNA?
Chapter 8
Success Criteria
 At the end of class today, you will be able to:
 Explain what a restriction enzyme is and what role they
play in the cell
 Explain how restriction enzymes have been used by
biologists as a tool in the manipulation of DNA
Biotechnology
biotechnology – manipulation of _______________
To study the functions of individual genes, molecular
biologists will cut them out of a genome and place
them into bacteria.
Basic Techniques
Six basic techniques will be covered:
1.
2.
3.
4.
5.
6.
DNA isolation
restriction enzyme digest
PCR
gel electrophoresis
sequencing
gene cloning
Isolating DNA
Before DNA can be manipulated, it needs to be
isolated from the cells.
1. Cell membranes are:
1. DNA precipitation
2. DNA isolation / storage
DNA Isolation
pp 366 - 367
Restriction Enzymes
Restriction Enzymes
restriction endonucleases –
recognition site –
Restriction Enzymes
Why are these enzymes used?
Where do they come from?
What role do they have in nature?
Restriction Enzymes
Recognition sites.
Palindromic.
Which of the following would be palindromic?
a)
b)
c)
d)
ATACTC
AACGTACGTT
CGGCCGGC
ACGCGT
Restriction Enzymes
Restriction enzymes are named according to the
organism from which it was identified.
Ex. EcoRI
E
co
R
I
- Escherichia
- coli
- strain RY13
- 1st enzyme in this strain
Restriction Enzymes
sticky ends –
If the following piece of DNA is cut with Xho1, what
ends would be created? (See page 367 for
recognition site)
5’-ATTCGCTACTCGAGCTCGATCCTCGAGGCGCAATC-3’
Restriction Enzyme Cutting
blunt ends –
DNA Ligase
T4 DNA ligase –
Recombinant DNA
recombinant DNA:
Recombinant DNA Animation