LAB 5: Getting the 16S rRNA genes from the Soil Genomic DNA in
the E. coli clones ready to send away for DNA sequencing
Preparing your clones to send away for sequencing analysis of
your 16S rRNA gene When you examine your transformation plates
after an overnight incubation, you should see hundreds of well
isolated colonies. In theory, each of them should contain the
vector plasmid with an insert of the 16s rRNA gene from one of
your soil sample bacteria. Since the vector plasmid contains a
kanamycin resistance gene, kanamycin resistance is a selectable
marker. The genetically engineered strain of E. coli that we
transformed is sensitive to kanamycin UNLESS it is expressing
the kanamycin resistance gene on the plasmid. E. coli that did
not take up a cloning vector plasmid and express its genes do
not form colonies on media with kanamycin. Kanamycin is an
antimicrobial compound that disrupts bacterial protein synthesis
and kill the cells. We know each of the vector plasmids in the
transformed E. coli growing o