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Questions and Answers
What is the primary purpose of using flanking primers with restriction sites in PCR recombination?
What is the primary purpose of using flanking primers with restriction sites in PCR recombination?
To generate overlapping sequences and allow for merging of two DNA fragments
How do you ensure that the PCR amplification of the two fragments results in a full-length product?
How do you ensure that the PCR amplification of the two fragments results in a full-length product?
By melting the DNA strands, allowing one in a thousand times for the fragments to anneal, and then extending the product
What is the role of the green and yellow parts in the PCR recombination process?
What is the role of the green and yellow parts in the PCR recombination process?
They are added to the respective fragments to generate overlapping sequences, allowing for the merging of the two fragments
What is the purpose of using the same restriction enzymes in the PCR recombination process?
What is the purpose of using the same restriction enzymes in the PCR recombination process?
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What is the final step in the PCR recombination process, after amplifying the full-length product using flanking primers?
What is the final step in the PCR recombination process, after amplifying the full-length product using flanking primers?
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