BIOC 300D2 Lecture Notes - Lecture 8: Chemotherapy, Coding Region, Provirus
Document Summary
Note that mrna is not that abundant in cells (1- Ribosomal and transfer rna are the first and second most abundant rna in cells, respectively. Regulation of gene expression can occur at 4 levels: transcriptional, post-transcriptional, translational, post-translational. A common technique to examine gene expression is to measure the level of individual mrnas (quantification) Techniques to monitor gene expression at the mrna level: northern blot: still used for particular applications such as the size of a mrna (need to run on gel) or track processing of mirna precursor. Run rna samples on an agarose gel, and set up transfer pyramid (transfer rna on gel to solid support, nitrocellulose). Add radio-active probe to nitrocellulose membrane, hybridization and expose membrane. Intensity of bands correspond to abundance of rna in sample. => time-consuming, use of radiation: rnase protection assay: mix sample rna with radioactive probe (complementary to target, single-stranded), and made them anneal together.