As C5-6 Technologies advances its biomass platform, Lucigen continues to expand its molecular biology products for gene cloning and genomics. Since the introduction of its first product, CloneSmart®, Lucigen has released other cloning products, including ClonePlex™-AK dual insert cloning kits, CopyRight™ large insert cloning kits, and BigEasy® linear cloning system.
Recently Lucigen discovered methods to clone genes and entire genomes directly from a few or just a single isolated cell. This capability, called PicoClone™, represents a billion-fold improvement in cloning compared to standard methods, says Dr. Mead. PicoClone makes it possible to clone genes from microbes that cannot be grown in the laboratory, such as extremophiles living in boiling hot springs. Lucigen researchers use PicoClone to discover superior types of enzymes for basic and biomedical research, molecular diagnostics, drug discovery, and industrial processes.
The first product developed using PicoClone is Pyrophage 3173 DNA Polymerase™, the first replicase cloned from a bacteriophage living in a boiling hot spring in Yellowstone National Park. Because PyroPhage 3173 contains a true DNA replicase, rather than the DNA polymerases found in all other current PCR enzymes, it gives more robust and truer replications, notes Dr. Mead. The thermostable properties of Pyrophage 3173 DNA make it ideal for thermocycling methods like PCR, he adds.
At the end of 2007, Lucigen released ExCyto PCR™, a new concept in DNA cloning and PCR analysis. ExCyto PCR incorporates a thermostable DNA polymerase into the E. coli chromosome, so there is no need for external DNA polymerases. “It’s a clever new way to bypass the work of purifying a gene and buying expensive DNA polymerases,” says Dr. Mead. ExCyto PCR allows researchers to insert a gene, incubate overnight, select a colony, and add cells to provided primers.
The ExCyto PCR kit comes with competent cells and all reagents for high efficiency ligation, transformation, and PCR. Highly reliable and reproducible amplification inserts of up to 5 kb are directly obtained from a colony sample. The PCR reagents provided are sufficient to perform at least 100 50 µL amplifications.
In June 2007, Lucigen and C5-6 Technologies were named commercial partners in the DOE’s Great Lakes Bioenergy Research Center (GLBRC). Based at the University of Wisconsin and funded with a five-year, $125 million grant, GLBRC includes investigators at Michigan State University, University of Florida, and Oak Ridge National Laboratory who are searching for ways to generate biofuels from biomass. By applying their enzyme technologies, Lucigen and C5-6 Technologies will contribute to expanding commercial opportunities for cellulosic biomass.