A Level Biology: Pyrosequencing

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How many types of activated nucleotides are used in a pyrosequencing reaction?

1

What are the 2 substrates used in pyrosequencing?

APS and luciferin

Give the 4 activated nucleotides used one group at a time in pyrosequencing:

ATP, TTP, CTP, GTP

What are the 4 enzymes used in pyrosequencing?

DNA polymerase, ATP sulfurylase, luciferase, apyrase

Explain how the pyrosequencing reaction produces photons of light:

a compilatory base pair is added to the single strand of DNA using an activated nucleotide and DNA polymerase which produces PPi as a result. PPi is converted to ATP by ATP sulfurase in the presence of APS. The ATP reacts with luciferin in the presence of luciferase to produce oxyluciferin which produces photons of light

What is high-throughput sequencing?

a variety of fast, cheap methods to sequence genomes

What does APS stand for?

adenosine 5' phophosulfate

What is the role of the DNA polymerase in pyrosequencing?

it catalyses the addition of a complimentary base pair from an activated nucleotide

What is the role of luciferase in pyrosequencing?

it catalyses the reaction between ATP and luciferin to form oxyluciferin

What is the role of ATP sulfurase in pyrosequencing?

it converts PPi to ATP in the presence of APS

What is the role of PPi in pyrosequencing?

it is converted to ATP by ATP sulfurase in the presence of APS

What is the role of APS in pyrosequencing?

it must be present for ATP sulfurase to catalyse PPi to ATP

What is the role of oxyluciferin in pyrosequencing?

it produces photons of light

What is the role of luciferin in pyrosequencing?

it reacts with ATP in the presence of luciferase to produce oxyluciferin

What is the role of ATP in pyrosequencing?

it reacts with luciferin in the presence of luciferase to produce oxyluciferin

What is the role of apyrase in pyrosequencing?

it removes left over bases from the reaction

Give an example of high throughput sequencing:

pyrosequencing

Give all the ingredients used in pyrosequencing:

single stranded DNA, primer, 4 enzymes, 2 substrates, 4 activated nucleotides

How does pyrosequencing methods compute the photons of light produced?

the light has different frequencies and single, double and triple peaks can form


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