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Hortense makes up a reaction mix to do Sanger sequencing on a lobster. She mixes the following together: lobster DNA, primers, four DNA nucleotides (A, C, G, and T), and DNA polymerase. The reaction does not work. What was the error in Hortense's reaction?

User EmptyStack
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2 Answers

7 votes
She didn’t cycle the reaction! She has to cycle as follows:
1. 95C-to denature the double stranded DNA
2. 50-55C depends on her primers- to anneal the primers to the single stranded DNA
3. 37-42C depending on the enzyme used - DNA polymerase extends from the primers to make the new DNA.

Also, are the dNTPs special? Either radioactive or with a fluorescence to measure/detect them?
User Ililit
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4 votes

Answer:

The answer is ( She should have added a small amount of a modified version of one of the four nucleotide.) Just took the test got it right.

Hope this helps:)

Step-by-step explanation:

User Keith Bloom
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