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A plasmid preparation is a method of DNA extraction and purification for plasmid DNA. It is an important step in many molecular biology experiments and is essential for the successful use of plasmids in research and biotechnology. [1] [2] Many methods have been developed to purify plasmid DNA from bacteria.
Plasmid Cloning Guides Molecular Cloning Guides —References to help scientists design plasmid cloning experiments, including tutorials on restriction enzyme digestion and PCR-based cloning. Molecular Cloning Protocols —Specific protocols for a variety of plasmid cloning techniques, such as isolation of bacterial colonies, DNA purification ...
Topoisomerase-based cloning (TOPO cloning) is a molecular biology technique in which DNA fragments are cloned into specific vectors without the requirement for DNA ligases.Taq polymerase has a nontemplate-dependent terminal transferase activity that adds a single deoxyadenosine (A) to the 3'-end of the PCR products.
The fertility plasmid or F-plasmid was discovered by Esther Lederberg and encodes information for the biosynthesis of sex pilus to aid in bacterial conjugation. Conjugation involves using the sex pilus to form a bridge between two bacteria cells; this bridge allows the F+ cell to transfer a single-stranded copy of the plasmid so that both cells contain a copy of the plasmid.
An origin of transfer – A plasmid with no origin of transfer is non-mobilizable. [ 2 ] The transfer genes – Though a functioning set of tra genes is necessary for plasmid transfer, they may be located in a variety of places including the plasmid in question, another plasmid in the same host cell, or even in the bacterial genome.
Plasmid-mediated resistance is the transfer of antibiotic resistance genes which are carried on plasmids. [1] Plasmids possess mechanisms that ensure their independent replication as well as those that regulate their replication number and guarantee stable inheritance during cell division.
A plasmid is a double stranded circular DNA molecule commonly used for molecular cloning. Plasmids are generally 2 to 4 kilobase-pairs (kb) in length and are capable of carrying inserts up to 15kb. Plasmids contain an origin of replication allowing them to replicate inside a bacterium independently of the host chromosome.
Recombinant DNA (rDNA) molecules are DNA molecules formed by laboratory methods of genetic recombination (such as molecular cloning) that bring together genetic material from multiple sources, creating sequences that would not otherwise be found in the genome.