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MBS1002 Biomedical Approaches

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Question:

What is the method of next generation sequencing (NGS)?

Author: Mirte Schattorie



Answer:

NGS is also called second generation sequencing. 1. isolate DNA and generate smaller fragments. 2. DNA fragments are coupled to different adaptors on both ends. 3. DNA fragment strands are then denatured. 4. separate DNA strands then bind with their adaptors to complementary sequences on the flow cell. 5. DNA is then amplified through PCR (while on the flow cell). 6. DNA is then denatured again and the strand that is not attached to the flow cell is washed away. 7. bridges are formed through the binding of the second adaptor to the cell. 8. bridges are amplified and both strands will stick to the flow cell. 9. bridge formation and bridge amplification is repeated several time to generate many copies. 10. once amplification is done, the reverse strand is cleaved and sequencing can start (the primer binds to the adaptors). 11. fluorescently labelled nucleotides are added and when the complementary nucleotide binds, it is excited by a laser and the fluorescent signal is obtained and the nucleotide is identified.


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