Introduction to sequencing

Question 1

Explain first generation of sequencing: Name, procedure and output

Answer 1

Sanger:

• fragment DNA
• clone it into plasmids and amplify
• add DNA polymerase, a primer and tagged nts (+ some ddNTPs to terminate chain)
• run capillary/gel and read code

Low output, long reads of high quality

Question 2

Give examples of 2nd generation sequencing

Answer 2

454, IonTorrent (both not used), BGISeq (has problems)

Illumina

Question 3

Give examples of use of NGS in the clinic

Answer 3

Diagnosis
Focused treatment (e.g. cancer and bacterial infections)

Question 4

Give examples of use of NGS in general

Answer 4

Metagenomics, ancient genomics and population genomics, RNA seq, single cell sequencing, genomic epidemiology…
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