Recombinant DNA Technology
A plasmid

All of these
usually contains one or more restriction sites
is a circular DNA molecule
always contains an origin of replication

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Recombinant DNA Technology
Knockout mice are created by

creating a chimera by fusing cells from two different cell lines
infecting the mouse with a retrovirus
mutagenizing a mouse and selecting for mutant offspring
transfecting embryonic stem cells with an altered gene sequence

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Recombinant DNA Technology
Some genetic diseases cannot be diagnosed by changes in restriction sites. Some of these can be detected by allele-specific oligonucleotide probes. These are

copies of the gene with an altered sequence so that a restriction site is inserted
short sequences that will hybridize only to a specific base sequence
PCR-amplified variable numbers of tandem repeats (VNTRs)
mutagenized copies of a gene

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