Question 1. You are sequencing a 350bp lizard gene using the Sanger dideoxy sequ

Question

Question 1. You are sequencing a 350bp lizard gene using the Sanger dideoxy sequencing method. You have amplified the region using the primer 5'-ATCCGGGCG-3'. What will be the length of the lowest band in your sequencing gel?

(A) Cannot answer the question with the provided information

(B) 350

(C) 10

(D) 9

(E) 351

Question 3. Which of the following answer(s) are TRUE (select all that apply)?

- PCR results in exclusively the desired region to be amplified . - PCR primers must be outside the region of interest .

- PCR primers must be flanking the region of interest. - Sequencing primers must be flanking the region of interest.

-All of the nucleotides used in Sanger Sequencing would also be able to be used for a PCR reaction where you are trying to amplify a gene of interest .

- Sanger sequencing requires dNTPs.

- A standard PCR reaction requires ddNTPs

- Small DNA fragments on a gel would be closer to the wells on a gel than larger DNA fragments.

Also can you give a bit explanation with your anwers.

Explanation / Answer

Answer 1: (b) 10. The smallest sequence will be of 10 bp because the primer is of 9 bp then last bp remaining for the sequence will come just after the primer hence the length of the smallest fragment will be of 10 bp.

Answer 3: statements that are correct are:-

1. PCR results in exclusively the desired region to be amplified.

2. PCR primers must be flanking the region of interest.

3. Sanger sequencing requires dNTPs.

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