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What is recombinant DNA technology?
Recombinant DNA technology involves techniques for obtaining, amplifying, and manipulating specific DNA fragments.
What is genetic engineering?
Genetic engineering is the process of producing modified DNA in vitro and introducing recombinant DNA into host organisms.
What is genomics?
Genomics is the cloning and molecular characterization of entire genomes.
What are restriction enzymes?
Restriction enzymes, or restriction endonucleases, are enzymes that cut DNA at specific sequences, creating fragments with sticky or blunt ends.
What is a restriction map?
A restriction map shows the locations of restriction enzyme sites within a DNA fragment, useful for DNA analysis and cloning.
What are cloning vectors?
Cloning vectors are DNA molecules used to carry foreign DNA into host cells for replication and expression.
What are the key features of a plasmid vector?
Plasmid vectors are small, circular DNA molecules with an origin of replication, polylinker, selectable marker, and reporter gene.
What is transformation in cloning?
Transformation is the process of introducing recombinant DNA into host cells, such as bacteria.
Who won the Nobel Prize in Chemistry in 1980?
Paul Berg won the Nobel Prize in Chemistry for his studies on recombinant DNA, particularly in the metabolism of galactose in E. coli.
What is a genomic library?
A genomic library is a collection of DNA fragments representing an organism's entire genome, each fragment cloned into a vector.
How many clones are needed for a human genomic library?
Approximately 100,000 clones are needed to represent the human genome in a BAC vector with 150 kb inserts.
What is a cDNA library?
A cDNA library is a collection of DNA fragments synthesized from mRNA, representing only the expressed genes in a specific tissue.
What is the main advantage of a genomic library?
Genomic libraries represent all regions of DNA equally, showing the intact genome structure in the cloned fragments.
What is the main advantage of a cDNA library?
cDNA libraries reveal the genes expressed in specific tissues, showing which mRNA is used for protein synthesis.
What is Southern blot analysis?
Southern blot is a method to detect specific DNA sequences in DNA samples using restriction enzymes and hybridization.
What are sticky and blunt ends in DNA?
Sticky ends have overhanging nucleotides, while blunt ends are straight cuts
both are products of restriction enzyme digestion.
What is a selectable marker?
A selectable marker is a gene introduced into a plasmid vector to identify and select cells that have taken up the plasmid.
What is a polylinker?
A polylinker, or multiple cloning site, is a short DNA sequence in vectors with multiple unique restriction enzyme sites for cloning.
What is a reporter gene?
A reporter gene in a vector allows researchers to visually confirm the expression or presence of the recombinant DNA in host cells.
What is ligation in recombinant DNA?
Ligation is the process of joining DNA fragments, such as inserting donor DNA into a vector, using enzymes like DNA ligase.
What is a hybridization probe?
A probe is a labeled single-stranded DNA or RNA sequence used to detect complementary sequences in DNA or RNA samples.
What is PCR?
Polymerase Chain Reaction (PCR) is a technique used to amplify specific DNA sequences, creating millions of copies.
What are BACs used for?
Bacterial Artificial Chromosomes (BACs) are vectors used to clone large DNA fragments in genomic studies, often for creating genomic libraries.
What are sticky ends used for in cloning?
Sticky ends allow DNA fragments to bind easily to complementary sequences in other DNA fragments, aiding in cloning.
What is a genomic equivalent?
A genomic equivalent is the number of clones needed to cover an entire genome once in a library.
What is a restriction digest?
A restriction digest uses restriction enzymes to cut DNA at specific sites, generating fragments for cloning or analysis.
What is transformation efficiency?
Transformation efficiency refers to the success rate of introducing foreign DNA into host cells during genetic engineering.
What is colony hybridization?
Colony hybridization is a technique to identify bacterial colonies with specific DNA sequences by hybridizing a labeled probe.
What is the purpose of creating a genomic library?
Genomic libraries enable researchers to study specific genes or entire genomes by providing access to cloned DNA fragments.
What is gel electrophoresis?
Gel electrophoresis is a technique to separate DNA fragments by size, using an electric current to move them through a gel matrix.
What is a vector's origin of replication?
The origin of replication is a DNA sequence allowing a vector to replicate within a host cell independently.
What is a plasmid?
A plasmid is a small, circular DNA molecule commonly used as a vector for cloning in bacterial cells.
What is a cDNA clone?
A cDNA clone is a DNA fragment synthesized from mRNA, representing only expressed genes in the library’s source tissue.
What are blunt ends used for in cloning?
Blunt ends can be joined without regard to sequence, but they often require additional steps to ensure stable ligation.
What is EcoRI?
EcoRI is a commonly used restriction enzyme that cuts DNA at the sequence GAATTC, creating sticky ends.
What is BamHI?
BamHI is a restriction enzyme that cuts DNA at the sequence GGATCC, also creating sticky ends for cloning.
What is the purpose of screening a library?
Screening identifies clones that contain a DNA sequence of interest, often using probes or selectable markers.
What is transformation in bacteria?
Transformation is the process where bacteria take up foreign DNA from their environment, often used in cloning.
What is DNA ligase?
DNA ligase is an enzyme that joins DNA fragments together by forming phosphodiester bonds.
What is ethidium bromide used for?
Ethidium bromide is a fluorescent dye used to stain DNA in gels, allowing visualization under UV light.
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