cloning of foreign DNA into plasmid

목차

1. Introduction
2. Background
3. Materials
4. Methods
5. References

본문내용

Introduction
Cloning DNA in Plasmid
- By fragmenting DNA of any origin (human, animal, or plant) and inserting it in the DNA of rapidly reproducing foreign cells, billions of copies of a single gene or DNA segment can be produced in a very short time. DNA to be cloned is inserted into a plasmid (a small, self- replicating circular molecule of DNA) that is separate from chromosomal DNA. When the recombinant plasmid is introduced into bacteria, the newly inserted segment will be replicated along with the rest of the plasmid.

Background
1. Units of ligase activity
- Weiss unit: One Weiss unit is defined as the amount of enzyme required to catalyze the exchange of 1 nmole of 32P from pyrophosphate to ATP, into Norit-adsorbable material in 20 minutes at 37℃.
- NEB unit: One unit is defined as the amount of enzyme required to give 50% ligation of Hind III fragments of λ DNA (5´ DNA termini concentration of 0.12 µM, 300 µg/ml) in a total reaction volume of 20 μl in 30 minutes at 16°C in 1X T4 DNA Ligase Reaction Buffer.
- 1 weiss unit is approximately equivalent to 60 cohesive end unit (=NEB unit)

참고 자료

http://www.ornl.gov/sci/techresources/Human_Genome/publicat/primer/fig11a.html
Sambrook, Russell. Molecular cloning (the 3rd edition). CSHL press. vol.1, 1.19-24
Sambrook, Russell. Molecular cloning (the 3rd edition). CSHL press. vol.1, 1.84-87
Sambrook, Russell. Molecular cloning (the 3rd edition). CSHL press. vol.3, A8.30-8.31