title: Bacillus cereus ATCC 14579 amplicon-based 12k microarray
status: Public on Mar 27 2008
contact.city: Wageningen
contact.first: Menno
contact.middle: van der
contact.last: Voort
contact.address: Bomenweg 2
contact.department: Agrotechnology and Food Sciences Group
contact.country: Netherlands
contact.institute: Wageningen UR
organism: Bacillus cereus ATCC 14579
manufacture_protocol: Amplicon based DNA-microarrays were constructed for B. cereus ATCC 14579 as described for Lactococcus lactis IL1403 (Kuipers et al., 2002; van Hijum et al., 2005) with modifications as detailed below. Amplicons were designed on 5199 genes selected from the 5311 annotated genes (ORFs smaller than 80 bp were omitted) on the genome of B. cereus ATCC 14579 (Ivanova et al., 2003). To reduce cross-hybridization between probe and target DNA sequences the amplicons had sizes of 70 – 700 bp (depending on gene sizes) and comprised the most unique part of a gene. The amplicons were synthesized by EuroGentec (Seraing, Belgium) in two amplification steps. In the first amplification step, primers were used with a unique tag-sequence for B. cereus ATCC 14579 (forward primers were extended with the sequence: 5’-TCGGGCAGCTGCTCC-3’; and reverse primers with the sequence: 5’-TGGCGCCCCTAGATG -3’). Two copies of each amplicon were present per array, resulting in microarrays comprising 10398 spots.