Probes design:
The complete genome sequence of Lactococcus lactis a Gram positive lactic acid bacterium has been published (A. Bolotin et al Genome Research 11: 731-753; 2001) and the nucleotide sequence is available from NCBI with accession number no: AE005176.
The genome contains 2310 ORFs with an average length of 900 bp. There are 370 paralogous families, comprising 1189 gene products, in the L. lactis genome (A. Bolotin et al; 2001). The MicroArrays were designed and developed in close collaboration with the Molecular Genetics group at the University of Groningen, The Netherlands (Prof. Dr. O.P. Kuipers, Dr. J. Kok).
The number of ORFs decreases to 1920 when duplicate genes and IS elements were removed from the list. 1920 pairs of primers were designed and synthesized to generate the L. lactis amplicons-based MicroArray.
We followed the two-step methods to generate C6-amino modified PCR products. The immobilized probes consist of 200-800 bp spotted in duplicate. The MicroArray contains the Lucidea Universal ScoreCard Controls, which is a set of artificial genes that function as universal references that are consistent from experiment to experiment and from species to species.
Quality control :
The L. Lactis MicroArrays are certified to generate a high quality data. All printed slides are previously selected for coating uniformity and low intrinsic background. The MicroArrays are visually inspected for spotting quality (doughnuts), uniformity, integrity and for perfect lignment of spots and grids. From each batch of printed MicroArray, three are randomly chosen for further quality control. We use the Terminal Transferase reaction to assess the amount of DNA covalently attached to the slides. Finally, we perform a real hybridization using labeled cDNA obtained from heat-choc cultured cells.
Legal notices
The use of this Array-related product in relation to the manufacture or use of nucleic acid arrays may be covered by the following patent owned by Oxford Gene Technology Limited ("OGT"): EP 0,373,203. The purchase of this product does not confer on the purchaser any rights or licences under any OGT patent. To enquire about a licence under OGT's patents, please contact licensing@ogt.co.uk.