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Recombinant Dna Technology: Ecori
Recombinant Dna Technology: Ecori
Recombinant DNA - DNA molecule containing DNA fragments (from different sources)
that are not naturally found
Vectors
Fragments produced after restriction enzymes cannot easily enter host cell
Gene carrier; also known as cloning vehicle
Characteristics of a Vector
It must be small, and unlikely to degrade during purification
It has markers (such as antibiotic resistance) that can indicate (in culture)
whether transformation has been successful
It must have an origin of replication so that DNA can be replicated by the host
cell's machinery
It has have several unique restriction sites so that the vector DNA will be cut
only in the desired location, and that several such locations will be available
for insertion of foreign DNA
These three steps correspond to one PCR cycle. This will be repeated, each
time doubling the number of copies of the DNA strand in the mixture. After 30 PCR
cycles, over one million copies of DNA strand can be made.
Applications of PCR
1. For rapid diagnosis of infectious diseases
2. Detection of rare pathological events such as mutation leading to cancer
3. Examine DNA of extinct species (mammoth, dinosaurs)
4. In criminal investigations, DNA fingerprints are prepared from the cells in a
tiny speck of dried blood or semen or at the base of a single human hair
5. Physicians can detect genetic defects in very early embryos by collecting a
sloughed off cells and amplifying DNA