BMS 607 Lecture Notes - Lecture 3: Agarose, Tandem Repeat, Real-Time Polymerase Chain Reaction
16 Dec 2016
School
Department
Course
Professor
Document Summary
Snp, indel, simple sequence repeat/microsatellite, cnv (very big), complex variant. Snps account for the vast majority of total sequence variation between humans genomes. ~ 18 million human snps have been identified. Pcr and cut: amplify gene-specific area -then cut with restriction enzymes. You need to know what you"re looking for- gene specific. Blot lights up from denatured single stranded dna that was transferred to gel that combines with dna with probe. After hybridization of probe to the blot, autoradiography reveals fragments in restriction digests that have sequences complementary to the probe. Restriction site-altering snps can be detected by southern blots. Pcr is shorter- 2 hours (amplify the area and cut and run) Alelle-specific oligonucleotide (aso)- specific for only one allele of the dna being tested. If there is a mismatch between probe and target, hybrid will not be stable at high temperature. Need to know wild type gene sequence.
