Question

In: Biology

1. In a paragraph, what are the major steps performed during next generation sequencing of DNA?...


1. In a paragraph, what are the major steps performed during next generation sequencing of DNA? How do these steps differ in the Illumina platform as compared to the Roche 454 platform?

2. In a paragraph, describe the major steps performed during DNA sequencing by the Pacific Biosciences, as well as the Oxford Nanopore Sequencing Technologies.

3. In a paragraph, describe the major advantages and disadvantages of each DNA sequencing platform.

Solutions

Expert Solution

Next generation sequencing of DNA have major three steps which are given below:-

  1. Library Preparation
    1. Input DNA fragment using enzymes and sonication methods
    2. Fragmentation of DNA having blunt end or overhangs
    3. END repairing and ligation using adaptor
    4. Fragment library preparation using specific adaptor
  2. Clonal Amplification
    1. Using Emulsion PCR method.
    2. Bridge amplification method.
  3. Sequencing
    1. Using Pyrosequencing
    2. Sequencing by ligation method
    3. Reversible dye incorporation

Roche 454 platform follow the pyrosequencing method on pyrosequencer technology while lllumina platform uses bridged technology for amplification.

Illumina yields the larger and correct contigs which are short in length in comparison to Roche 454. Illumina technology is cheapest technology in comparison to Roche 454, which is costly. Roche 454 has major advantage of resolving the palindrome sequence and repetitive sequence.

Comparison of All sequencing techniques:-


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