PCL102H1 Lecture Notes - Lecture 2: Dna Microarray, Rad51, Dna Extraction
Document Summary
It binds ahre, dre, or xre (response elements) and recruits coactivators upregulats cyp1a1. Recognized sequence: tngngtg: lollipop box, estrogen receptor (from pcl302, epigenetics, methylation of cpg islands represses gene expression, environmental factors can affect epigenetics, histone tails can be modified in many ways (e. g. phosphorylation, acetylation, methylation, etc. ) A type of immunoprecipitation experimental technique used to investigate the interaction between proteins and dna in the cell. It aims to determine whether specific proteins are associated with specific genomic regions, such as transcription factors on promoters . Establish protein interactions with in vivo dna sequences. Allows study of histone modifications on transcription in vivo: methodology, treating cells or tissues briefly with formaldehyde. Formaldehyde crosslinks dna and protein (cytosine, lysine, arginine). Cell nuclei are isolated (though whole cell extracts work too: fragmenting the dna/protein complexes. Ideal fragment size is 600 (200: bp. More sonication = less smearing of bands. Samples are centrifuged and supernatant (chromatin) is isolated.
