Bioinformatics 13: High throughput sequencing 1

Question 1

What is capillary electrophoresis and why is useful?

Answer 1

Analytical technique that separates ions based on their electrophoretic mobility.

Electrophoretic mobility is dependent upon charge, viscosity, and the atom’s radius

Automated, high-throughput, accurate,

Question 2

High-throughput applications of Sanger sequencing?

Answer 2

Whole Genome Shotgun (WGS) for genome sequencing

Re-sequencing for variation discovery - SNPs etc.

De novo ‘end sequencing’ Structural Variants (SVs)

Question 3

What is ‘massively parallel’ sequencing? Main companies/machines involved?

Answer 3

Use of clone amplification to make millions of copies of single template molecules

Physically immobilise clusters of template molecules

• monitor reactions optically via CCD camera
• decode sequence by computational analysis of snapshots
• map short reads to reference genome

Major players

• > [454] (1.4mil reads per experiment)
  - > Ion torrent (80 mil reads per exp.)

-> Illumina (20bil reads per exp.)