Plasmid Isolation

CytoGenomics offers small- and large-scale preparations of high quality plasmid DNA for all molecular biology and gene therapy applications. Cytogenomics offers yields of up to 100 mg. Automated instrumentation is used to perform rapid plasmid miniprep DNA templates for sequencing in the 96 well format. A variety of other prep services for BAC, PAC, cosmid and lambda clones are also offered

 

Cloning and subcloning

CytoGenomics uses standard restriction enzyme, PCR, and ligation technology to clone or subclone known genes. RT-PCR can be applied to clone cDNAs of interest from supplied RNA, or RNA can be purified by Cytogenomics. 5’-RACE-PCR can be performed to identify and clone the 5’ end of a cDNA. 3’RACE can be applied if the 3’ end is unknown. Genes can be recloned to construct full length clones.

 

Metabolic Engineering

CytoGenomics will design appropriate metabolic systems in vector of your choice to get optimal metabolic functions. Functions are directed by the client we find appropriate Metabolic solutions in the vector. We combine pathways to give you a solutions to your problem.

CytoGenomics will design your protein for optimal expression in E. coli, yeast, or baculoviral systems. Proteins can be designed with “HIS-tags”, “GST-tags”, or other system of your choice. Cytogenomics recommends a regulatable expression system to provide optimum protein yields, but will work with the vector system of your selection. Expressed proteins are analyzed by SDS-PAGE, purified using the affinity tags or other chromatographic step, and verified by N-terminal sequence analysis, if requested. Standard proteins are provided at ~85% purity. Additional purification steps are available at your discretion.

 

Gene Synthesis

CytoGenomics has applied sophisticated technologies to construct completely synthetic “designer genes” expressing the exact desired protein. Genes are synthesized by combining overlapping complementary oligonucleotides, filling in the gaps with a thermostable nucleotide polymerase, and amplifying the product by PCR. These amplified products are cloned into the desired plasmid vector and overexpressed. Genes up to ~2.0 kbp have been synthesized, but there is no theoretical limit to the size.

Synthetic “designer genes” normally have unique DNA sequences that have been modified to optimize the codon usage for the target host. In addition, restriction sites, leader sequences, homing signals, or other modified sequences can be incorporated for special applications.

 

Site Directed Mutagenesis

CytoGenomics normally performs site-directed mutagenesis using modifications of a mega-primer PCR protocol. Using this technology, substitutions of 1-10 bp can easily be generated. These substitutions are useful for any site-directed mutagenesis project, and have been often applied for alanine-scanning type of protein functional probing or “linker-scanning” analyses. Clones can be verified by sequence analysis.

Custom Array development

We provide custom array development for RNA expression studies that are modified to suit customer needs. We use Incyte clones & where required from other sources.

• Mini Array of sizes of upto 200 genes
• Maxi Array of sizes upto 8000 genes
• Complete Array & Tissue specific Arrays