DNA sequencing and personalised medicine

Question 1

Sanger sequencing: principle, requirements and limitations

Answer 1

locate specific SNP and so determine sequence; required lot of DNA, fluorescent or radioactive dd primer; 1 sequence added at a time

Question 2

next-generation sequencing: method and requirements

Answer 2

PCR amplification then all clusters sequenced at same time OR single molecule pulled through tiny channel as each nucleotide has different shape; reference sequence needed

Question 3

next-generation sequencing: advantages

Answer 3

faster, cheaper to sequence whole genome

Question 4

next-generation sequencing: disadvantages

Answer 4

less accurate, short read length

Question 5

next-generation sequencing: purpose

Answer 5

target specific genes, coding sequence, protein-bound regions, epigenome (DNA methylation sites)