Lab 3: Continue Soil Microbial Communities & Diversity Project
Today you will continue your identification of the total
bacteria in your soil habitat by measuring the concentration of
the genomic DNA that you isolated in Lab 2. When your DNA is at
the proper concentration (100ng/μL) you will amplify, in a
polymerase chain reaction (pcr), part of the 16S rRNA gene from
hundreds or thousands of the different species of bacteria (some
of which are culturable and most of which are non-culturable)
using "universal" bacterial primers and a proof-reading DNA
polymerase. If you are unfamiliar with the basics theory behind
pcr amplification of target DNA, please review it in at the
following web site: The bacterial primers we will use are called
27 Forward and 1492 Reverse. The numbers refer to positions on
the gene where these single strand primers will anneal and where
the DNA polymerase will begin to use your soil isolates' genomic
DNA as templates to create double stranded sections of the 16s
rRNA gene from the va