19, Below is the description on the DNA sequencing by pyrosequencing. Fill in th

Question

19, Below is the description on the DNA sequencing by pyrosequencing. Fill in the bl ank. (10pt) "Pyrosequencing technique is based on sequencing by-synthesis principle and on the detection of released uctbehi's during DNA synthesis. It employs a series of four enzymes to accurately detect nucleic acid sequences during the synthesis. In Pyrosequencing the sequencing primer is hybridized to a single-stranded DNA biotin-labeled template and mixed with the enzymes; DNA polymerase, ATP sulfurylase, luciferase and apyrase, and the substrates adenosine 5' phosphosulfate (APS) and luciferin. Cycles of four polymerization reaction in which inorganic polymerase. Subsequently the released are separately added to the reaction mixture iteratively. The cascade starts with a nucleic acicd is released as a result of nucleotide incorporation by by to oxyluciferin in is quantitatively converted to in the presence of , which drives the conversion of ,producing visible light in amounts that are proportional to the amount of ATPs. The light is then detected by a photon detection device such as a charge coupled device (CCD) camera or photomultiplier ATP and non-incorporated dNTPs in the reaction mixture. There is a certain time interval (usually 65 seconds) between each nucleotide dispensation to allow complete degradation. For this reason, dNTP addition is performed one at a time. Because the added nucleotide is known, the sequence of the template can be determined". is a nucleotide-degrading enzyme, which continuously degrades

Explanation / Answer

There are 10 blanks

1. Pyrophosphate

2. dNTP (De-oxynucleotide triphosphate)

3. Pyrophosphate

4. Pyrophosphate

5. ATP

6. ATP sulfurase

7. APS

8. Luciferin

9. Luciferase

10. Apyrase

Their is nothing to describe about those blanks because these all based on facts (that which enzyme convert which substrate into which product)

You have to memorize those enzyme names and works and uses.

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