title: cDNA microarray Trymanosoma cruzi - v.1
status: Public on Mar 04 2005
contact.city: São Paulo
contact.first: Ricardo
contact.last: Vêncio
contact.email: rvencio@vision.ime.usp.br
contact.state: SP
contact.address: Av. Prof. Lineu Prestes, 748 - Butantã
contact.department: Departamento de Bioquímica
contact.country: Brazil
contact.web_link: http://www.vision.ime.usp.br/~rvencio/Tcruzi/
contact.institute: Instituto de Química da Universidade de São Paulo
contact.phone: +55 (11) 3091-3839
organism: Trypanosoma cruzi
description: DNA microarrays were constructed with 710 ESTs obtained from non-normalised and normalised cDNA libraries of CL Brener epimastigotes . Polymerase chain reaction (PCR) amplification of the ESTs (average length 800 bp) was obtained with T3 and T7 primers using DNA polymerase (BIOLASE). The amplification products were purified with Multiscreen plates (Millipore). Inserts representing 20 cloned T. cruzi coding and non-coding genes , control sheared DNAs and oligonucleotides  were also immobilised on the glass slides with Generation III Microarray System spotter (Molecular Dynamics) according to manufacturer's instructions. The DNA probes were distributed in 10 sub-arrays, containing 288 spots each. Each sub-array was spotted in duplicate (left and right side of the slide) resulting in at least 6 and at most 60 replicates depending on probe. Detailed spotting map is available at supplemental web-site. After immobilisation, DNA was UV-irradiated at 254 nm (Stratagene lamp) with 50 mJ total energy.