title: UTU Chicken BursaEST 37K array
status: Public on Jul 19 2010
contact.city: Stockholm
contact.first: Pekka
contact.middle: Juhani
contact.last: Kohonen
contact.address: Nobelsväg 13, Box 210
contact.department: IMM Institute of Environmental Medicine
contact.country: Sweden
contact.institute: Karolinska Institutet
organism: Gallus gallus
manufacture_protocol: BursaEST array (from the dkfz426 cDNA library) BursalEST library construction: The bursas of 2-wk-old chicken of the inbred CB strain were excised and a cell suspension was prepared. The chicken bursa consists mainly of B cells but Ficoll gradient centrifugation was used to remove contaminating epithilial and red blood cells to achieve a B cell content of about 95% B cells. Poly-dT reverse transcribed mRNA was ligated to a linker and cloned into pSPORT1 plasmid. Average insert length was about 1.3 kb. Library is housed at RZPD (Deutsches Ressourcenzentrum für Genomforschung), Heidelberg, Germany as Chicken Bursa cDNA (number 426, or dkfz426). Sequences are submitted in GenBank under accession AJ392050-AJ3994559. The library is not normalized. Spotting the array: Plasmid DNA was grown up and PCR amplified using primers to the linker sequences at RZPD. PCR products were purified and spotted to a nylon filter macroarray in duplicate within a 4x4 subarray. The array contains 36864 (192x192) features including 14592 cDNA clones (in duplicate), spike control spots (not used in hybridizations) and empty spots. redundancy.The array can be hybridized several times and quality of spots on the array can be assesed by hybridizing with linker oligos to determine DNA content of a spot.