BIOT 732: Genetic Engineering II - Cloning Techniques

A technique using PCR to introduce mutations into a DNA sequence by designing primers with desired changes.

A method for introducing mutations into a plasmid using a PCR reaction with primers carrying the desired mutation.

A cloning technique that uses a special vector containing a topoisomerase enzyme to directly ligate PCR products.

A method that uses a mix of enzymes to efficiently assemble multiple DNA fragments, regardless of length or end compatibility, in a single-tube isothermal reaction.

A type of cloning based on site-specific recombination, using enzymes from bacteriophage lambda to transfer DNA segments.

A specialized type of Gateway cloning that allows for simultaneous assembly of multiple DNA fragments into a single destination vector.

A one-pot, one-step cloning method that uses Type IIS restriction enzymes to assemble DNA fragments.

Artificial endonucleases designed to target specific DNA sequences and cleave DNA at those sites.

A type of DNA-binding domain that recognizes specific DNA sequences and can be engineered to target specific regions for manipulation.

A variation of ZFNs that uses a different DNA-binding domain derived from bacterial transcription activator-like effectors (TALEs).

Proteins derived from bacteria that bind to DNA and can be engineered to target specific DNA sequences.

A cloning technique that uses PCR to amplify a fragment of interest with added attB sites, which are then used for recombination.

The second step in Gateway cloning, where a DNA fragment is transferred from an Entry vector to a Destination vector.

A plasmid vector that contains the attL sites, flanking the gene of interest, created through the BP reaction.

A vector that contains attR sites, used to receive the gene of interest from an Entry clone in the LR reaction.

A suicide gene that prevents the growth of bacteria if it is not removed.

A feature that eliminates undesired restriction sites from a destination vector, ensuring that only the intended sites are used for cloning.

Type IIS restriction enzymes used in Golden Gate Assembly, recognizing and cutting DNA outside their recognition sequences.

The unique four-base pair overhangs generated by Type IIS restriction enzymes during Golden Gate assembly.

A type of cloning that uses a donor vector containing attB sites outside a specific DNA fragment, which is then recombined into an Entry clone.

A plasmid vector that contains specific attB sites that are targeted during the BP reaction in Gateway cloning.

A technique for introducing mutations into a plasmid by using PCR with mutagenic primers and subsequently digesting the parental DNA.

A method for introducing mutations that involves altering the DNA sequence by introducing a single base change.

A technique used to remove specific restriction sites from a DNA sequence, commonly used to avoid unwanted cuts during Golden Gate Assembly or other cloning methods.

A specialized set of plasmids designed for efficient cloning of DNA fragments through recombination reactions, using attB sites.

A method for creating custom-designed transcription activator-like effector nucleases (TALENs) for gene editing, utilizing Golden Gate Assembly.

A powerful genome editing tool that involves fusion of transcription activator-like effectors (TALEs) to a FokI enzyme.

Artificial nucleases that can target specific DNA sequences for cutting, useful for gene editing and manipulation.