Multiple Choice
What is the enzyme used to anneal the sticky ends of a plasmid to an inserted gene in a cloning reaction?
3. Acids and Bases
Organic Chemistry Reactions
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What occurs during the annealing stage of a PCR reaction?
A
Primers bind to the single-stranded DNA template.
B
The DNA strands are separated by heating.
C
The DNA polymerase extends the DNA strand.
D
The reaction mixture is cooled to stop the reaction.
Verified step by step guidance1
Understand that PCR (Polymerase Chain Reaction) is a technique used to amplify a segment of DNA, making millions of copies of a particular DNA sequence.
Recognize that PCR involves three main stages: denaturation, annealing, and extension.
During the annealing stage, the temperature of the reaction is lowered to allow primers to bind to the single-stranded DNA template. This is crucial for the specificity of the PCR, as primers are short sequences of nucleotides that are complementary to the target DNA region.
The annealing temperature is typically set a few degrees below the melting temperature (Tm) of the primers, ensuring that they bind specifically to the target sequence without forming non-specific interactions.
Once the primers are bound, the DNA polymerase can extend the DNA strand in the next stage of PCR, using the primers as starting points for DNA synthesis.
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