BIOM30002 Lecture Notes - Lecture 20: Restriction Fragment Length Polymorphism, Fluorescent Tag, Polyacrylamide Gel Electrophoresis
Document Summary
High frequency of f508del in aus from migrants. Use cftr gene sequence to design primers that flank mutation. Polyacrylamide gel used because agarose can"t resolve a small 3bp mutation. N/n: thickened band indicates homozygous; cf/cf: homozygous for f508del (3bp shorter); n/cd: heterozygous. This test will only pick up the f508del mutation. N/cf could be a carrier of f508del or be a compound heterozygote. F508del first mutation that was tested for diagnostically. Testing typically used pcr +/- restriction enzymes (rflps - restriction fragment length polymorphisms) Multiplex pcr with multiple primers that can detect several mutations. Number of mutations in testing panels differs throughout the world - constantly evolving. Some mutations are specific to particular ethnic groups. So different countries use different panels to reflect ethnic groups. Previously, in victoria a total of 12 mutations were tested in the initial diagnostic panel, together accounting for ~80% of mutations. A further 7 mutations were tested in the extended panel.
