Page 77 - Clinical Biochemistry

PCR-RFLP

• Restriction Fragment Length Polymorphism
• Cutting a DNA sequence using restriction enzymes into pieces → specific enzymes cut
specific places

Figure 47: Restriction Fragment Length Polymorphism

Starting DNA sequence:
5’-TAATTTCCGTTAGTTCAAGCGTTAGGACC
3’-ATTAAAGGCAATCAAGTTCGCAATAATGG

Enzyme X Enzyme X
5’-TTC- 5’-TTC-
3”-AAG- 3”-AAG-

5’-TAATTT 5’-CCGTTAGTT 5’-CAAGCGTTAGGACC
3’-ATTAAA 3’-GGCAATCAA 3’-GTTCGCAATAATGG

212

The Technology: Genotyping

• Uses a microarray to measure a limited predefined set of SNPs

• Very high throughput (fast)

• Cheap

• Excellent coverage of common variation (up to 5,000,000 SNPs)

Figure 48: microarray