Clemson University Genomics Institute

The BRC has extensive experience in creating custom large insert BAC libraries with an average insert size generally ranging between 120 and 160kb. CUGI has produced numerous plant, animal and microbial BAC libraries encompassing many different taxa in the tree of life. BAC libraries at CUGI are generally constructed using one or more of the following restriction enzymes; HindIII, BstyI, EcoRI, BamHI, and/or MboI. The current charges for producing a BAC Library is $0.30 per clone for academic and non-profit entities and $0.50 for commercial organizations. This rate includes all library construction costs, picking and arraying the library into 384-well microtitre plates to produce the master copy, and one replicate or working copy of the library. The plates will be bar-coded and placed in cardboard freezer storage boxes that hold 36 plates per box. Filters and additional library copies can be produced at our standard rates.

High quality random-sheared shotgun libraries of BAC clones and whole genomes are regularly produced in the BRC. The libraries have been used extensively in DNA sequencing projects throughout the world. Average insert sizes are 3-5kb and can be constructed up to 10kb. The libraries are generally picked and arrayed into 96-well microtitre plates amenable to most high-throughput DNA sequencing production systems. Please contact for a quote.

The newest type of library to be produced at the Clemson University Genomics Institute are Fosmid libraries. FOSMIDs are commonly used for preparing genomic libraries when a smaller insert size is desired. The inserts have an average size of 40 kb and are produced by random shearing, yielding a more uniform coverage of the genome than other library types. FOSMIDs are excellent candidates for closing gaps in a whole genome sequence and physical maps due to their uniform coverage. Additionally, they are popular for metagenomic and expression studies. The entire 40 kb insert can be shuttled into an expression vector more easily than a larger insert and the higher copy number per cell allows for easier manipulation and cheaper downstream applications. Current costs for fosmid library construction are $0.15/clone.

The BRC produces high quality cDNA libraries. The libraries are well-suited for EST sequencing and other gene discovery applications. The cDNA libraries are generally cloned into a standard high-copy plasmid vector and picked and arrayed into 96-well microtitre plates amenable to most high-throughput DNA sequencing production systems. The current charge for cDNA library construction for academic and non-profit entities is $7,500.00 per library which includes library construction and one copy of the arrayed library up to 192 plates (18,432 clones). The charge for commercial entities is $10,000 per library. The recipient will also receive the unused portion of the ligation. The customer must isolate and supply the RNA for cDNA preparation to CUGI. In addition, the cDNA libraries we produce are not normalized.

The BRC is capable of picking as many as 40,000 colonies per day with its two Genetix Q-bots. The colonies require plating on Genetix Q-Trays at a density of 2000-3000 total colonies per tray. The machines are capable of blue/white color selection as with IPTG and X-Gal or black/white selection based on the new Sigma X-al. The current cost is $0.08 per colony for academic and non-profit entities and $0.12 for commercial organizations which includes Q-plate preparation, media, bar-coded label and freezer storage boxes for the plates.

The BRC can replicate library plates at the rate of $12.00 per microtitre plate for academic and non-profit entities and $25.00 for commercial organizations. This includes the plate, media, bar-coded label and freezer storage boxes for the plates. The center can replicate over 250 master plates into 2 individual replicas per day.

The BRC re-arrays libraries robotically using a selected list of clones which are entered into the Q-Bot's computer and then selectively picked from the original library plates and placed in new microtitre plates to create a new re-arrayed version of the library. This can be done in both 96 and 384-well formats. Charges are project specific and quotes can be obtained by contacting the BRC for more information.

The BRC grids libraries from microtitre plates onto Hybond N+ membrane (filters) in high-density colony filter arrays using Genetix Q-Bots. The current production rate is a maximum of 90 filters per day. In most cases, 18,432 individual colonies are arrayed in a 4x4 pattern in duplicate (double-spotted), for a total of 36,864 spots per filter. Higher density arrays can be provided following consultation with the BRC. Please contact us for more information. Each filter is stamped with a library name and a unique serial number. The current cost for producing filters is $150.00 per filter for academic and non-profit entities and $250.00 for commercial organizations. For custom hybridization service quotes using BAC filters please also contact the BRC.

Libraries may be depostied at CUGI for public distribution to commercial entities and public instituitions. Clones are sold at standard CUGI prices. Two copies of the library will be made at standard microtitre plate copying charges to provide one copy to actively distribute clones from and another copy that will be saved to produce additional future copies. We have two options available in regards to storage charges. For the first option, a fee of $100.00 per freezer box (a freezer box will hold up to 36 plates) is charged per year for storage. If the library is to be stored and distributed indefinitely, then -80C freezer space can be purchased based on library size and the current cost of the -80C freezers. If this second option is desired, please contact the BRC for a quote.