NUSCTX 170 Lecture Notes - Lecture 17: Isopropyl Alcohol, Chloroform, Hydrogen Bond
3 Dec 2020
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Lab 17: did mtch2 knockdown affect gene expression. Overview of the flow of information from dna to protein in a eukaryote: both coding and noncoding regions of dna transcrived to mrna, some regions are removed (introns) during initial mrna processing. This causes all protein to immediately denature and precipitate and inactivates degradation enzymes such as cellular rnases. Phenol disrupts cell membranes: after addition of chloroform and vigorus mixing, phase separation occurs, the top colorless aqueous phase contains rna. Intermediate white solid phase contains genomic dna: bottom pink organic phase contains protein, when the top phase is separated and mixed with isopropanol, rna precipitates. Guanidium thiocyanate: chaotropic salt, efficiently precipitates proteins and nucleic acids by disrupting non-covelent interactions inside these macromolecules. Interferes with hydrogen bonding network: by this mechanism, it also acts as dnase and rnase inactivator during the process of.
