Bacterial Genomic DNA isolation practical ( Phenol-chloroform Method)
I have seen many people (including me) struggle to find a good practical protocol with principle, roles of reagents , observations and discussions all clubbed together as one. The literature is always dispersed throughout net across various papers and documentation. Therefore here I'm providing a summarized short but to the point practical for the Phenol-Chloroform Method of genomic DNA isolation from Bacteria(say E.coli). ------------------------------------------------------------------------------------------------------------------------------ AIM Isolation of genomic DNA from bacterial cells MATERIALS REQUIRED LB medium ( per ml, 10 mg tryptone, 5 mg yeast extract, and 5 or 10 mg NaCl), Incubator Reagents 20% SDS , Phenol: chloroform: isoamylalcohol (25:24:1) , RNase , NaCl solution (5M) , chilled ethanol(70%) , TE(Tris-EDTA) buffer[10mM Tris-Cl(pH8.0), 1mM EDTA(pH 8.0] , Lysis buffer[TE buffer, proteinase K, NaCl/SD...