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1 Jan 2018

In dideoxy sequencing method,only one primer is used in each reaction tube during DNA synthesis. Why not use both forward and reverse primers in the same tube simultaneously, as in PCR?

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4 Jan 2018
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Related questions

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?

2. Which of the following answer(s) are true (select all that apply)?

Question options:

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.

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 2. True or False

If cell has a mutated gyrase, it means that it will be unable to properly join together okazaki fragments and therefore will result in many fragmented pieces of DNA.

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.

2. This question concerns PCR and DNA sequencing. a. Describe what would happen in a PCR experiment if you only used one primer instead of two primers. b. Suppose you perform PCR using only one copy of a double-stranded DNA molecule. How many copies of this double-stranded DNA molecule would be present after 15 cycles? c. We usually think of enzymes being the most active at around 37?C (near body temperature). In PCR, however, DNA synthesis occurs at 72?C during each cycle. What is unique about the DNA polymerase that is used in PCR? d. Dideoxynucleotides (ddNTPs) play an important role in DNA sequencing. What is the structural difference between a nucleotide (NTP), a deoxynucleotide (dNTP), and a dideoxynucleotide (ddNTP).

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