title: VUMC MACF human 5K BAC v16
status: Public on Aug 30 2006
contact.city: Amsterdam
contact.first: Daoud
contact.last: Sie
contact.email: d.sie@vumc.nl
contact.address: De Boelelaan 1117
contact.department: Pathology
contact.country: Netherlands
contact.institute: Vrije Universiteit Medical Center
contact.phone: +31 20 4442428
organism: Homo sapiens
description: - clones were mapped to the UCSC July 2003 Freeze of the Human Golden Path (NCBI build 34)
  - oncobacs were mapped using their overgo-primers and are available via www.ensembl.org
manufacture_protocol: as described in: Carvalho et al (J. Clin. Path.,2004, vol. 57, 644-646). DNA from the 1 Mb resolution Sanger BAC clone set (see Ensembl), the Caltech OncoBac set, and in house clones of interest, supplemented with clones from the Children’s Hospital Oakland Research Institute (CHORI), amounting to a total of 4678 clones with known chromosomal positions, was isolated according to the CHORI protocol. Amplification of the DNA was done by ligation mediated polymerase chain reaction (PCR) according to Snijders et al. (Nat. Genet.,2001, vol.29, 263-264) and PCR clones were purified using Montage PCR µ96 filter plates (Millipore BV, Amsterdam, the Netherlands). Clones were spotted in triplicate at a concentration of 1 µg/µl, in 150 mM sodium phosphate (pH 8.5), on CodeLink™ slides (Amersham BioSciences, Roosendaal, the Netherlands), using a SpotArray 72 robot (Perkin Elmer Life Sciences, Zaventum, Belgium) equipped with SMP3 pins (TeleChem International, Sunnyvale, CA, USA). After printing slides were processed and blocked prior to use according to the manufacturer?s protocol.