title: Penn/CHOP Homo sapiens 4.5K (BAC array for studying Neuroblastoma)
status: Public on Feb 28 2007
contact.city: Philadelphia
contact.first: Jayanti
contact.last: Jagannathan
contact.email: jagannathan@email.chop.edu
contact.state: PA
contact.address: 34th & Civic center blvd
contact.department: Oncology
contact.country: USA
contact.institute: Childrens Hospital of Philadelphia
contact.phone: 2674262286
organism: Homo sapiens
manufacture_protocol: BAC clone starter cultures were grown in 700ul 2X YT broth with 12.5 ug/mL chloramphenicol at 37 deg C for ~15h. Full cultures of 2.5 mL broth were inoculated from starter cultures and grown for 15 h in 48-well blocks. DNA was extracted using QAGEN real prep kits using high throuhput 96 well blocks. BAC DNA was then amplified using degenerate oligonucliotide primers (Cheung and Nelson 1996). Each BAC was amplified twice at different annealing temperatures (58 and 60 deg C). PCR products were pooled and purified with 96 well QiaQuick kits. Each clone DNA was lyphilized and resuspended in 50% DMSO/water. A minimum of two replicates per clone were printed on each slide using the aqueous DMSO buffer as spotting solution. Arrays were printed on Corning CMT Ultra GAP slides.