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Next Generation Sequencing
Helicos – True Single Molecule Sequencing (tSMS)
Library Preparation
• Shearing of DNA
• Sonication
• Nebulization
• Enzyme digestion
• Denature dsDNA
• Tailing
• Blocking
Sample Loading and Flow Cell
Preparation
• Sample Fragments Loaded
• Nucleotides are added sequentially
Sequencing by Synthesis
Work Cited
• Ambry Genetics. Making Sense of NextGent Sequencing. Kelly Gonzalez, MS,
CGC, and Senior Manager of Clinical Genomics. Available at
http://www.ambrygen.com/sites/default/files/pdfs/NERRG_4-10-
12_Making_Sense_of_NetGen_Sequencing_KG(3).pdf. Access verified May
21, 2014.
• Omixon. Allen Van Deynze, 2010; Solanaceae Coordinated Agricultural Project.
Next Generation Sequencing. Available at http://www.omixon.com/the-basics-
of-next-generation-sequencing/. Access verified May 21, 2014.
• http://en.wikipedia.org/wiki/Helicos_single_molecule_fluorescent_sequencing
Access verified July 30, 2014.
• Seqll. Howe it Works. 2014. http://seqll.com/the-sample-process/ Access
verified August 1, 2014.
Recommended Video
Helicos True Single Molecule Sequencing
https://www.youtube.com/watch?v=_-
MK3MGfPvY
Next Generation Sequencing
Single Molecule Real Time Method