cpn60 based metagenomics
cpn60 UT PCR can be used to
generate amplicon libraries from complex microbial communities.
Examples based on both clone library analysis and direct
pyrosequencing have been published. To date, pyrosequencing has
been done on the Roche 454 platform (both FLX and Titanium).A complete list of multiplexing ID (MID) tagged primers used by our group to date is available here. Please note that we have optimized our metagenomic protocols to exploit a "primer cocktail" approach we developed previously (Hill et al. 2006) to improve representation of microbial diversity in metagenomic libraries. Thus, the MID-tagged primers are used in sets of four (2 forward, 2 reverse).
A detailed protocol for application of cpn60 UT PCR to profiling complex microbial communities, including library production and data analysis, was published in Methods in Molecular Biology in 2011.
Updated 2011-06-07