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Gene Cloning

Gene Cloning

Creating a copy of an organism or molecule from another entity

Restriction Enzymes

DNA-cutting enzymes that cleave phosphodiester bonds in DNA strands

Plasmid DNA

Circular, self-replicating DNA found in bacteria, used to carry and clone other DNA fragments

Recombinant DNA Technology

Technology that allows for the cutting and joining of DNA fragments, and insertion into a plasmid

Bacteriophage vectors

clone DNA fragments up to 15 kb; infect the bacterial species

Restriction Site

Specific sequence of bases in DNA recognized and cut by restriction enzymes

Palindrome

Sequence that reads the same forward and backward on opposite strands of DNA

Sticky Ends

Overhanging single-stranded ends of DNA fragments created by certain restriction enzymes

Blunt Ends

Double-stranded ends of DNA fragments created by certain restriction enzymes

Recombinant DNA

DNA formed by joining together DNA fragments from different sources

Transformation

Process of inserting foreign DNA into bacterial cells

Antibiotic Selection

Process of selecting transformed bacteria by growing them on plates with different antibiotics

Blue-White Selection

Method to identify recombinant bacteria based on the color of colonies on a plate

Dna Libraries

Collections of cloned DNA fragments from a particular organism

Genomic DNA Libraries

Libraries containing fragments of an organism's entire genome

cDNA Libraries

Libraries containing complementary DNA synthesized from mRNA

Polymerase Chain Reaction (PCR)

Technique for amplifying specific DNA sequences in a short period of time

Taq DNA Polymerase

DNA polymerase isolated from Thermus aquaticus, used in PCR due to its heat stability

Cloning PCR Products

Rapid and effective method of cloning DNA fragments amplified by PCR

PCR primers

Primers are created to match the beginning and end sequence of certain sections

These are short DNA sections that set the boundaries for what will be copied

DNA probes are

short, single-stranded segments of DNA used in DNA hybridization experiments.

multiple cloning site (MCS)

site that can be recognized by multiple restriction enzymes

cloning vector

DNA molecules that can carry foreign DNA into a host cell and replicate there.

gel electrophoresis process

enzymes are used to chop DNA from sample into fragments, fragments are then placed into wells; electrical current is run through the gel and fragments are attracted depending on charge; smallest/lightest/most charged move far, thus creating banding pattern used to identify and compare

DNA ligase

enzyme that chemically links DNA fragments together

bacterial artificial chromosome (BAC)

A large plasmid that acts as a bacterial chromosome and can carry inserts of 100,000 to 300,000 base pairs.