CSB350H1 Study Guide - Final Guide: Agrobacterium Tumefaciens, Arabidopsis Thaliana, Tris
15 Jul 2018
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Arabidopsis thaliana from salk line: contain dna plasmid from argobacterium tumefaciens (bacteria) with t-dna + antibiotic resistance. Argobacterium tumefaciens vacuum infiltrated into a. thaliana flowers seeds t1 has plasmid if survive antibiotic. Lab using seeds from mutant plant with t-dna insertion in chromosome 3, at locus atskl1. Reagents liq. nitrogen breaks cell wall of plant cells, helps access dna material and inner contents. Ctab denatures proteins on dna (histones, associated proteins) Tris is a buffer, maintains neutral ph (7-8) Dnaase with mg will be active, will damage dna unless edta chelates with mg, stops dnase. Tbe = tris + boric acid + edta. Part 1: plant dna extraction liq. nitrogen + plant material grind to fine powder. Part 1: making agarose gel for electrophoresis of pcr products. 4 students, 1 agarose gel (make depending on size/position of expected pcr band) see how many wells you need use dna ladder to help semi-quantify later.
