Sequencing

What generation do real-time sequencers belong to?

• 1st generation

• 3rd generation

• Next Generation Sequencing

This image represents which high throughput sequencing method

• Sanger sequencing

• Nanopore

• PacBio

• Illumina

·         What sequencing technology is represented in the image?

• Illumina

• 454

• PacBio

• Nanopore

·         Imagine that you are in an expedition across the Arctic Ocean and that you need to find the bacterial diversity in a sample you just obtained. Which sequencing technology is more advisable to use and why?

• Illumina, because it brings the most reliable results.

• Nanopore, because minion is portable and easy to use outside the lab.

• Nanopore, because it is the only technology that allows us to find bacterial diversity.

• PacBio, because it is always better to use this technology.

·         The main advantage of third generation sequencing regarding next-generation sequencing is…

• Less preparation and sequencing time.

• More accuracy of the sequences.

• Starting material is the single-molecule, it doesn´t require prior PCR.

• Cheaper technologies.

·         What considerations must we keep in mind while loading our flowcell?

• Avoid introducing air bubbles into the flow cell.

• Close priming port and spot on port before starting the run.

• Remove air from the flowcell before adding the priming mix.

• All answers are correct.

·         What is the purpose of the 16S PCR performed in this protocol?

• Amplification of a small region of 16S gene.

• Amplification of the whole 16S gene, addition of the corresponding index and the first adaptors needed to sequence.

• Amplification of all sample’s DNA.