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ServicesAt MetGen, we improve your target proteins properties by high throughput screening. Provide us with the target enzyme sequence and we will improve the desired properties by massive focused mutagenesis and high throughput screening. We perform all steps from site specific mutagenesis design, combinatorial DNA libraries construction to library screening and recombinant enzyme assay with substrate of choice.
1. Designing Design of specific multi site directed mutagenesis in the amino acid sequence of the desired protein for construction of combinatorial DNA library. Using MetGen technology combinatorial library design narrows mutagenesis to the targeted regions of the protein. This significantly decreases the complexity of the library and reduces the number of clones for screening. 2. Construction One of the most popular approaches to create diversity in the protein of interest is oligonucleotide-directed mutagenesis, which introduces variations at specific sites or short stretches of the corresponding gene resulting in altered protein sequences. An alternative to the site-directed mutagenesis is random mutagenesis. However, random mutagenesis creates a huge number of irrelevant mutations. MetGen technology implements a special library construction method Smart Codon Permutator, which creates a library containing all combinations from predefined sets of aminoacids at predefined positions. 3. Cloning Cloning of DNA library or cDNA library into an expression plasmid to produces cDNA expression library in target organism. Combinatorial DNA library cloning can be performed in various organisms (e.g. E.coli, yeast, phage ). We guarantee over 95% clones containing insert in our libraries. 4. Picking & Growing 
Clone selection by automated colony picking with colony picker (QPix2) and processing them in 96- or 384 well format. At MetGen we have Genetix QPix2, an extremely efficient and robust 96-pins picking robot with an extensive selection of uses. The QPix2 can pick up to 3000 bacterial or yeast clones per hour from various plate format and inoculates single colonies to 96- or 384 well plate. With Qpix we are able to do various applications like colony picking, library management and protein evolution etc.
5. Testing & Analyzing We have capability to analyze cell homogenates for desired activities with your target micro-organism or chosen substrates. For testing we have wide selection of tools such as HPLC, TLC, photometric, luminescence and absorption measurements and DNA sequencing of requested mutant clones, micro-organism DNA sequence. 6. Remixing Once favorable mutant clones for chosen substrate are found, clones will be sequenced and analyzed for sequence remixing. Sequences which contain useful mutations are mixed by means of MetGen unique method Sequence Blender for new screening round. |
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