title: MPIIB_Mtb_V2
status: Public on May 01 2007
contact.city: Berlin
contact.first: Hans-Joachim
contact.last: Mollenkopf
contact.email: mollenkopf@mpiib-berlin.mpg.de
contact.address: Charitéplatz 1
contact.country: Germany
contact.institute: Max-Planck-Institute for Infection Biology
contact.phone: +49 30 28460 482
organism: Mycobacterium tuberculosis
manufacture_protocol: Microarray capture probes were obtained as a collection of  “Array-Ready” 70mer DNA oligonucleotides at 600 pmol each comprising 4,325 M. tuberculosis-specific sequences and 25 controls (Operon Biotechnologies). The major portion of M. tuberculosis-specific oligonucleotides was designed from 3,924 predicted protein coding sequences of M. tuberculosis H37Rv (Sanger Center). A smaller portion was designed from the strain CDC-1551, sequenced and predicted by The Institute for Genomic Research (TIGR). At least 371 predicted genes from CDC-1551 were found to have less than 97% homology to M. tuberculosis H37Rv predicted genes, and therefore these sequences were included in the M. tuberculosis probe set. The oligonucleotides were suspended at a final concentration of 100 µM in Nexterion spot2 buffer (Schott) and printed by a BioRobotics MicroGrid II robot (Genomic Solutions) onto glass slides coated with poly-L-lysine at 50–55% humidity and room temperature using MicroSpot 2500 quill pins (Genomic Solutions). Oligonucleotides were cross-linked to the support by UV irradiation with a total energy of 240 mJ UV and blocked with succinic anhydride and 1-methyl-2-pyrrolidone in borate buffer (Fluka).