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Questions and Answers
What is the first step in Recombinant DNA technology?
What is the role of restriction enzymes in Recombinant DNA technology?
What is the purpose of ligation in DNA cloning?
What term describes the process of introducing recombinant DNA into a recipient host cell?
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What is the outcome of effectively transforming cells/organisms in Recombinant DNA technology?
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What does the term 'clone' refer to in DNA cloning?
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What is the technology used for producing artificial DNA by combining genetic materials from different sources?
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Who discovered restriction enzymes in the year 1968?
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What is the role of ligases in recombinant DNA technology?
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How do restriction endonucleases cut DNA?
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What is the purpose of DNA primers in the PCR process?
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What is the main function of Exonucleases in recombinant DNA technology?
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During which PCR machine step does denaturation of double-stranded DNA occur?
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What do sticky ends refer to in recombinant DNA technology?
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What enzyme aids in the synthesis of a complementary strand of DNA in the PCR process?
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What happens to the original DNA sample during the denaturation step in the PCR machine?
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Which step in the PCR process involves cooling the sample mixture to allow DNA primers and DNA polymerase to bind to individual strands of DNA?
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What is the function of the nucleotide solution mix in the PCR process?
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What is the term used to describe the process of forming a new complementary strand of DNA after the nucleotides pair with individual DNA strands?
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Which temperature range is involved in the cycling process of heating and cooling during DNA amplification?
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How many times is the cycling process of heating and cooling repeated using the thermal cycler?
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What happens to the resulting DNA sequence each time the heating/cooling cycle is conducted by the cycler?
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Why can millions of copies of DNA be formed after 35 doubling cycles?
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What makes the work of ligases easy while binding the desired gene to the vector?
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Which of the following is not a component of vectors used in recombinant DNA technology?
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What feature of plasmids and bacteriophages makes them popular as vectors in recombinant DNA technology?
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Which step in the process of recombinant DNA technology involves introducing the recombinant DNA into the host organism?
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What is the main role of the host organism in recombinant DNA technology?
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What is the sequence of nucleotides within a vector from where replication starts known as?
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Study Notes
Recombinant DNA Technology
- Also known as Genetic Engineering
- Involves combining different genetic materials (DNA) from different sources to produce artificial DNA
History of Recombinant DNA Technology
- Emerged with the discovery of restriction enzymes in 1968 by Swiss microbiologist Werner Arber
Tools of Recombinant DNA Technology
- Enzymes: restriction enzymes, polymerases, and ligases
- Restriction enzymes: cut the DNA at specific points, producing sticky ends
- Polymerases: synthesize DNA
- Ligases: bind DNA molecules
- Vectors: carry and integrate the desired gene into the host organism
- Plasmids and bacteriophages are commonly used vectors due to their high copy number
Process of Recombinant DNA Technology
- Step 1: Isolation of genetic material
- Step 2: Cutting the gene at recognition sites using restriction enzymes
- Step 3: Amplifying the gene copies through PCR
- Step 4: Ligation of DNA molecules
- Step 5: Insertion of recombinant DNA into the host
PCR (Polymerase Chain Reaction)
- A process to amplify a single copy of DNA into thousands to millions of copies
- Requires four components: DNA or RNA sample, DNA primers, DNA polymerase, and nucleotide solution mix
- PCR process has 4 steps: collection, preparation, amplification, and post-PCR clean-up
- PCR machine steps: denaturation, annealing, and extension
- Denaturation: heat breaks hydrogen bonds, separating DNA into single strands
- Annealing: cooling allows DNA primers and polymerase to bind to individual strands
- Extension: new complementary strand of DNA is formed, resulting in a new duplicate double-stranded DNA molecule
DNA Cloning
- A clone is a cluster of individual entities or cells that are descended from one progenitor
- Recombinant DNA is introduced into a recipient host cell, which gets multiplied and expressed in the form of the manufactured protein under optimal conditions
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Description
Learn about the key steps involved in Recombinant DNA technology, including isolation of genetic material, cutting the gene at recognition sites using restriction enzymes, and amplifying gene copies through Polymerase Chain Reaction (PCR).
