Biology
Cut the DNA fragment and plasmid using the same restriction enzyme. Add guanines to DNA fragment blunt ends and terminal transferase, to produce a single chain of guanines to both the blunt ends of the plasmid vector. Add cytosines to vector blunt ends, to produce a single chain of cytosines to both blunt ends of the DNA fragment.
Mix the cut plasmid and cut DNA fragment together for recombination to occur. Single chain guanine will bind to single chain cytosine by complementary base pairing. Add ATP and DNA ligase to seal the nicks by forming phosphodiester bonds in the sugar backbone.
Add recombinant DNA to bacteria culture for transformation to take place. Use heat shock and add calcium chloride to facilitate uptake of recombinant DNA.
From here on, answer is question specific...
Culture bacteria in agar plate containing ampicillin (assuming original plasmid had ampicillin resistant gene). Bacteria that has taken up the plasmid vector, whether reannealed or with the foreign DNA will be resistant to ampicillin and grow on the agar plate. ...
- Q: Compare And Contrast The Structure Of Collagen And Cellulose
The monomer of collagen is an amino acid whereas the monomer of cellulose is beta-glucose. The monomers in collagen are linked by peptide bonds whereas those of cellulose are beta (1,4)-glycosidic bonds. Collagen is an alpha helix polypeptide with a turn...
- Transcription
Each gene contains a specific nucleotide sequence called the promoter region and it contains the initiation site, where proteins, called transcription factors, and RNA polymerase enzyme recognize and bind to. The enzyme-DNA-protein complex then causes...
- Mammalian Satellites Consist Of Hierarchical Repeats
KEY CONCEPTS:Mouse satellite DNA has evolved by duplication and mutation of a short repeating unit to give a basic repeating unit of 234 bp in which the original half, quarter, and eighth repeats can be recognized.In the mammals, as typified by various...
- Individual Genomes Show Extensive Variation
KEY TERMS:Polymorphism (more fully genetic polymorphism) refers to the simultaneous occurrence in the population of genomes showing variations at a given position. The original definition applied to alleles producing different phenotypes. Now it is also...
- Restriction Endonucleases Are A Key Tool In Mapping Dna
KEY TERMS:Restriction endonucleases recognize specific short sequences of DNA and cleave the duplex (sometimes at target site, sometimes elsewhere, depending on type). A restriction map is a linear array of sites on DNA cleaved by various restriction...
Biology
Q: Explain how a foreign DNA fragment can be introduced into a plasmid that produces blunt ends.
Cut the DNA fragment and plasmid using the same restriction enzyme. Add guanines to DNA fragment blunt ends and terminal transferase, to produce a single chain of guanines to both the blunt ends of the plasmid vector. Add cytosines to vector blunt ends, to produce a single chain of cytosines to both blunt ends of the DNA fragment.
Mix the cut plasmid and cut DNA fragment together for recombination to occur. Single chain guanine will bind to single chain cytosine by complementary base pairing. Add ATP and DNA ligase to seal the nicks by forming phosphodiester bonds in the sugar backbone.
Add recombinant DNA to bacteria culture for transformation to take place. Use heat shock and add calcium chloride to facilitate uptake of recombinant DNA.
From here on, answer is question specific...
Culture bacteria in agar plate containing ampicillin (assuming original plasmid had ampicillin resistant gene). Bacteria that has taken up the plasmid vector, whether reannealed or with the foreign DNA will be resistant to ampicillin and grow on the agar plate. ...
- Q: Compare And Contrast The Structure Of Collagen And Cellulose
The monomer of collagen is an amino acid whereas the monomer of cellulose is beta-glucose. The monomers in collagen are linked by peptide bonds whereas those of cellulose are beta (1,4)-glycosidic bonds. Collagen is an alpha helix polypeptide with a turn...
- Transcription
Each gene contains a specific nucleotide sequence called the promoter region and it contains the initiation site, where proteins, called transcription factors, and RNA polymerase enzyme recognize and bind to. The enzyme-DNA-protein complex then causes...
- Mammalian Satellites Consist Of Hierarchical Repeats
KEY CONCEPTS:Mouse satellite DNA has evolved by duplication and mutation of a short repeating unit to give a basic repeating unit of 234 bp in which the original half, quarter, and eighth repeats can be recognized.In the mammals, as typified by various...
- Individual Genomes Show Extensive Variation
KEY TERMS:Polymorphism (more fully genetic polymorphism) refers to the simultaneous occurrence in the population of genomes showing variations at a given position. The original definition applied to alleles producing different phenotypes. Now it is also...
- Restriction Endonucleases Are A Key Tool In Mapping Dna
KEY TERMS:Restriction endonucleases recognize specific short sequences of DNA and cleave the duplex (sometimes at target site, sometimes elsewhere, depending on type). A restriction map is a linear array of sites on DNA cleaved by various restriction...