BIOL 205 Lecture Notes - Lecture 10: Har Gobind Khorana, Kary Mullis, Pyrococcus Furiosus
Document Summary
How to monitor mrna expression level for every gene: global transcriptional. Profiling: carry out thousands northern blots, instead dna microarrays were developed, dna microarrays for global transcriptional profiling were not feasible before the sequencing of whole genomes. Immobilized mrna population hybridized with labeled dna probe representing one or two genes. Immobilized dna probes representing all possible genes hybridized with labeled mrna population. Need to achieve two things: (i) immobilize (array) thousands of. Dna probes specific for each individual mrna gene product (ii) label mrna populations. The probes can be cdnas (~ 1kb) or oligonucleotides (20-70 mers) Hybridize to array wash monitor fluorescence for each fluorescent dye. Array of 1046 cdnas probed with fluorescently labelled cdna made from bone marrow mrna. Tells you whether you need new protein synthesis to change transcript level. Each row is a different gene, each column a different time point.
