Lecture 22

Question 1

Direct detection methods for hypothesis driven

Answer 1

• southern-RFLP, PCR-RFLP, PCR sizing, ASO blot
• FISH
• Northern, Western
• Sanger sequencing

Question 2

Methods query the entire genome

Answer 2

• karyotype
• microarray: CGH, SNP, cDNA
• SKY
• NGS(1 gene, related genes, exomes by WES, genome by WGS)

Question 3

Next Generation Sequencing

Answer 3

SsDNA fragments ligated with adaptors
Attached to a microarray chip, all sequenced at same time by a common primer
Data collected by laser and sequence analyzed by computer

Question 4

Read depth in NGS: different nucleotides at same position

Answer 4

• polymorphism
• a rare variant
• possible mutation the explains the condition in the patient