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Brassica rapa 24k Microarray
- Expression
profiling was conducted with the Brassica 24k Microarray. The 24k
Microarray was designed from 24,963 unigenes. Among these, the direction of
19,900 was known and 6 probes per gene were designed. The direction of
5,063 genes was not known and designed 6 probes for sense and anti-sense
direction, respectively. Probes were duplicated in 2 blocks. In total,
360,312 probes were designed. The average size of probe is 60-nt long with
adjusting its Tm value 75 to 85 °C. The microarray was manufctured at
NimbleGen inc (http://www.nimblegen.com/). Random GC probes (40,000) to
monitor the hybridization efficiency and four corner fiducial controls
(225) were included to assist with overlaying the grid on the image.
- To
assess the reproducibility of the microarray analysis, we repeated the
experiment two or three times with independently prepared total RNA. The
normal distribution of Cy3 intensities was tested by qqline. The data was
nomalized and processed with cubic spline normalization using quantiles to
adjust signal variations between chips and Rubust Multi-Chip Analysis (RMA)
using a median polish algorithm implemented in NimbleScan (Workman et al.,
2002. Genome Biol. 3: research0048.1 - research0048.16 ; Irizarry et al.,
2003. Nucleic Acids Research 3:e15).
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Hybridization
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Genes on
microarray
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