12 Questions
What ensures that each small DNA fragment fixed on the beads can be the unique template for amplification in the sequencing process?
Presence of complementary oligos on the beads
What is the purpose of using a PTP plate with special nanopores in the pyrosequencing process?
Fix the position of each bead for convenience in sequencing
What initiates the release of a pyrophosphate group in the pyrosequencing process?
Pairing of dNTP with template DNA
How does the pyrophosphate group produced during DNA synthesis lead to fluorescence in pyrosequencing?
Binds to ATP sulfuric acid chemical enzymes
What is the significance of using a CCD camera in the pyrosequencing process?
To record the signal of fluorescence from fluorescein molecules
Why does 454 sequencing technology conduct each reaction in an independent nanopore on the PTP board?
To reduce mutual interference and sequencing bias
What is the main principle of sequencing in the Roche 454 system?
Using a spray method to break DNA samples into small fragments and adding adapters at both ends
What distinguishes the DNA library construction process of Roche 454 from Illumina?
Using a spray method to break DNA samples into small fragments and adding adapters at both ends
What is the key technology behind emulsion PCR in the Roche 454 system?
Forming a large number of independent reaction spaces for DNA amplification
How does Roche 454 system ensure that each DNA template is amplified in an independent reaction space?
By fixing single stranded DNAs with 28um beads buried in emulsion
What is the role of mineral oil in the emulsion PCR process of Roche 454 system?
To form numerous small water droplets wrapped by it
Why is the use of emulsion PCR important in the Roche 454 system?
To amplify DNA templates in independent reaction spaces efficiently
Learn about the main principle of sequencing on the Roche 454 sequencing system, including the preparation of DNA library using a spray method to break DNA samples into fragments and the addition of adapters. Compare the library construction process with Illumina's method.
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