Using the principles of second generation sequencing technology we learned in class, describe in a step...

Using the principles of second generation sequencing technology we learned in class, describe in a step by step fashion how targeted sequencing of DNA and RNA were performed in this study? Write your response in paragraph form.

In general, be able to explain in a step by step fashion the workflows involved in carrying out sequencing with each of the different sequencing platforms utilized in these studies. Know what each step accomplishes. Write your response in paragraph form.

What are the advantages and disadvantages of each kind of platform (2nd, 3rd, 4th) generation)?

Expert Solution

Answer to first para:

Principle of second generation sequencing is similar to Sanger sequencing. To prepare the targeted nucleic acid (DNA/RNA) THE FOLLOWING STEPS ARE FOLLOWED.

1.Genomic DNA is isolated and randomly broken into fragments.

2.Template is created and amplified by Polymerase chain reaction

3.Single stranded template is immobilised on a solid support

4.The primer(RNA) molecule is covalently attached to the solid support and it is hybridised with template.DNA polymerase is made to bind to the immobilised primed template to initiate the second generation sequencing reaction.

5. It results in population of identical templates.Each one is taken for sequencing reaction.They aligned to a known reference sequence and data is analysed.

Answer to 3rd Para:

Advantages:

Second generation is most commomnly used,substancially cheaper. It is faster and helps in massive sequencing data.It is most commonly used.

In Third generation nucleici acids are sequenced directly and have higher output.

In fourthgeneration sequencing genomic analysis is directly in the cell(in vivo) and it is the most recent.

gladiator answered 1 year ago

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Using the principles of second generation sequencing technology we learned in class, describe in a step...

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