A. Transformation of ds DNA into cells
B. Denaturation and annealing of oligonucleotide primers with mutation
C. Repair of nicks in mutated plasmid inside cells
D. Extension and incorporation of primers by using DNA polymerase
E. Digestion of unmutated parental DNA template with methylation specific nuclease and annealing of newly synthesized strands
Choose the correct answer from the options given below :
Site-Directed Mutagenesis Steps Explained
To correctly order the steps of site-directed mutagenesis, we follow the logical progression of the molecular process:
- Denaturation and annealing of oligonucleotide primers with mutation (B): This is the crucial first step where the synthetic DNA primers, designed to contain the desired mutation, are hybridized to the template DNA.
- Extension and incorporation of primers by using DNA polymerase (D): A DNA polymerase enzyme extends the primers, synthesizing a new DNA strand complementary to the template. This incorporates the mutation into the newly synthesized strand, creating a heteroduplex molecule.
- Digestion of unmutated parental DNA template with methylation specific nuclease and annealing of newly synthesized strands (E): In many protocols, the original, non-mutated template DNA (often methylated) is selectively degraded using specific nucleases (like DpnI) to eliminate it and enrich the population of the desired mutated DNA.
- Transformation of ds DNA into cells (A): The processed, mutated DNA is then introduced into competent host cells (e.g., bacteria) via transformation.
- Repair of nicks in mutated plasmid inside cells (C): Once inside the host cell, the cellular DNA repair machinery acts on the plasmid DNA, sealing any nicks and ensuring the stable replication of the mutated plasmid.
Therefore, the correct sequence representing the process of site-directed mutagenesis is B, D, E, A, C.
Similar Questions
- Lectins are :
- Which of the following viruses has a segmented genome ?
- Which of the following heavy metal containing chemical agent is used for treatment of burns ?
- Blue-white screening is based on the activity of :
- Which growth phase is most important for antibiotic production ?
- What is Trichoderma harzianum used for in agriculture ?
- Which of the following is always present in a Virus ?
- The enormous diversity exhibited by MHC molecules within a species originates from :
- Selection for traits that are sensitive to population density and are favoured at high densities is known as :
- Aminopterin is added in HAT medium to :
Important Questions from Mixed Topic (CUET PG)
- Who was the founder of Bolshevik Communist party?
- What is the key guide to statecraft in the realist tradition?
- Chronologically arrange the events in the Cold War period.
A. Berlin Wall is constructed
B. Communist China joins the UN
C. Soviet invasion of Czechoslovakia
D. Berlin Blockade
Choose the correct answer from the options given below: Morgenthau's principles of political realism are:
A. Politics is rooted in permanent and unchanging human nature which is basically self centred, self-regarding and self-interested
B. Politics is an autonomous sphere of action and cannot therefore be reduced to morals
C. International Politics is an arena of conflicting self-interests
D. The ethics of international relations is situational ethics which is very different from private morality
Choose the correct answer from the options given below:Who among the following political thinkers consider the anarchical self help system to be a compelling factor for States to maximise their relative power positions?
