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A strip of eight PCR tubes, each containing a 100 μL reaction mixture Placing a strip of eight PCR tubes into a thermal cycler. The polymerase chain reaction (PCR) is a method widely used to make millions to billions of copies of a specific DNA sample rapidly, allowing scientists to amplify a very small sample of DNA (or a part of it) sufficiently to enable detailed study.
Polymerase cycling assembly (or PCA, also known as Assembly PCR) is a method for the assembly of large DNA oligonucleotides from shorter fragments. The process uses the same technology as PCR, but takes advantage of DNA hybridization and annealing as well as DNA polymerase to amplify a complete sequence of DNA in a precise order based on the single stranded oligonucleotides used in the process.
The first step in Gateway cloning is the preparation of a Gateway Entry clone. There are a few different ways to make entry clone. Gateway attB1 and attB2 sequences are added to the 5' and 3' end of a gene fragment, respectively, using gene-specific PCR primers and PCR amplification. The PCR amplification products are then mixed with a propriet
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 ...
Mixing this destination plasmid with the entry clones and the LR clonase mix will allow for recombination to occur between the attR and attL sites. A destination clone is produced, with the fragment of interest successfully inserted. The lethal gene is inserted into the original vector, and bacteria transformed with this plasmid will die.
Molecular cloning takes advantage of the fact that the chemical structure of DNA is fundamentally the same in all living organisms. Therefore, if any segment of DNA from any organism is inserted into a DNA segment containing the molecular sequences required for DNA replication, and the resulting recombinant DNA is introduced into the organism from which the replication sequences were obtained ...
Functional cloning is a molecular cloning technique that relies on prior knowledge of the encoded protein’s sequence or function for gene identification. [ 1 ] [ 2 ] [ 3 ] In this assay, a genomic or cDNA library is screened to identify the genetic sequence of a protein of interest.
The thermal cycler (also known as a thermocycler, PCR machine or DNA amplifier) is a laboratory apparatus most commonly used to amplify segments of DNA via the polymerase chain reaction (PCR). [1] Thermal cyclers may also be used in laboratories to facilitate other temperature-sensitive reactions, including restriction enzyme digestion or rapid ...
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