THE MOLECULAR BASIS OF INHERITANCE
RECOMBINANT DNA
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Question
[CLICK ON ANY CHOICE TO KNOW THE RIGHT ANSWER]
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Isolate recombinant DNA from bacteria
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Identify bacteria with recombinant DNA
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Identify cell with target DNA to be cut by the restriction enzyme
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Identify bacteria with plasmid to be used as a vector
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Detailed explanation-1: -A genetic screen or mutagenesis screen is an experimental technique used to identify and select for individuals who possess a phenotype of interest in a mutagenised population. The use of chromogenic substrate to detect a particular enzymatic activity is the basis to screen the desired clone.
Detailed explanation-2: -Today, the E. coli bacteriophage is one of the most widely used vectors used to carry recombinant DNA into bacterial cells. This virus makes an excellent vector because about one-third of its genome is considered nonessential, meaning that it can be removed and replaced by foreign DNA (i.e., the DNA being inserted).
Detailed explanation-3: -The most accurate way to verify your recombinant colonies is by Sanger sequencing. Plasmid DNA is first isolated from an overnight bacterial culture. Once completed, the insert can be identified using sequencing primers appropriate for the selected vector.
Detailed explanation-4: -There are six steps involved in rDNA technology. These are – isolating genetic material, restriction enzyme digestion, using PCR for amplification, ligation of DNA molecules, Inserting the recombinant DNA into a host, and isolation of recombinant cells.