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Sample GSM743646 Query DataSets for GSM743646
Status Public on Apr 01, 2012
Title 382128A10_Ki11_R2_T1
Sample type RNA
 
Source name Ki11, above ground seedling tissue
Organism Zea mays
Characteristics pooling type: pooled multiple seedlings
genotype: Ki11
classification: Tropical
Treatment protocol No treatments
Growth protocol Maize seeds were grown in a greenhouse with 50% field soil and 50% Metromix with a 16hour light and 8 hour dark treatment
Extracted molecule total RNA
Extraction protocol RNAs were isolated using the commercial TRIzol (Invitrogen, Carlsbad, CA; cat# 15596026) from above-ground tissue. RNAs were purified by Lithium Chloride treatment followed by 3M sodium acetate (0.1 vol) and 95% ethanol (2.5 vol) precipitation.
Label Cy3
Label protocol Purified RNAs (10ug per sample) were reverse transcribed and labeled according to the array manufacturer protocol (NimbleGen Arrays User’s Guide: Gene Expression Analysis v3.2).
 
Hybridization protocol Per sample, ~20μg of Cy3- or Cy5-labeled RNAs were hybridized for 16-20 hours at 42°C using the NimbleGen Hybridization System. Post hybridization, slides were washed (NimbleGen Wash Buffer Kit) and dried for two minutes by centrifugation.
Scan protocol Slides were immediately scanned using the GenePix 4000B Scanner (Molecular Devices, Sunnyvale, CA) according to the array manufacturer protocol.
Data processing Array images and data were processed using NimbleScan software. Briefly, images from each slide were separated into 12 subarrays and aligned to a grid to extract signal intensity for each feature on the array. Experimental integrity was verified by evaluation of the signal intensities of the sample tracking control features for each subarray. In addition, metrics reports were produced for each array to report the signal uniformity across the array and the intensity of known empty features, random probes and experimental probes. Signal-to-noise ratios were estimated by dividing the average signal intensity of the experimental gene probes (signal) by the average signal intensity of the random sequence control probes (noise). Only slides with a signal-to-noise ratio ≤ 2 were retained. NimbleScan was used to generate RMA-normalized (Irizarry et al. 2003) gene expression values from the spatially-corrected probe signal intensities on a per probe and per gene basis. Normalized gene expression values across multiple replications (technical or biological) of the same genotype were averaged, when possible. Comparisons of the distributions of signal intensity for the random sequence controls to the experimental gene probes on each array (Figure SM2) were used to determine a reasonable signal threshold for positive expression across all slides. The plots demonstrate that in most cases, the random sequence control log2 signal intensity is ≤ 10 suggesting that signals above that threshold are true signal. Hence, genes with average probe log2 signals of >10 in at least three arrays were retained as expressed (N=19,792 expressed genes). Since genetic polymorphisms could contribute to differences in hybridization between transcripts from some genotypes and probes developed based on the B73 reference sequence we further filtered the probeset based on a previous comparative genomic hybridization dataset developed using many of the same genotypes and the same array platform (Swanson-Wagner et al., 2010). Any probes that exhibit substantially reduced CGH values for at least three genotypes (26,937 probes) were removed resulting in a set of 46,167 probes that detect expression of 18,242 genes with one to four probes per gene. This dataset of N=18,242 expressed genes detected by probes without substantial evidence for polymorphisms was used for all analyses.
 
Submission date Jun 16, 2011
Last update date Apr 01, 2012
Contact name Nathan M Springer
E-mail(s) [email protected]
Phone 6126246241
Fax 6126251738
Organization name University of Minnesota
Department Plant Biology
Street address 1445 Gortner Ave
City Saint Paul
State/province MN
ZIP/Postal code 55108
Country USA
 
Platform ID GPL13736
Series (1)
GSE30036 Reshaping of the maize transcriptome by domestication

Data table header descriptions
ID_REF
VALUE Per gene expression values are derived from background-corrected RMA normalized data using NimbleScan (ver2.0) software. Each gene is represented by 1-4 probes that do not show significant comparative genomic hybridization variation.

Data table
ID_REF VALUE
chr10:100098205-100100917 9.8121
chr10:100123370-100125355 9.6017
chr10:100877502-100879751 11.9906
chr10:100903641-100921394 10.9536
chr10:10092665-10093736 5.5166
chr10:10134037-10138754 13.7437
chr10:101365191-101367444 11.2655
chr10:101535511-101540308 12.6555
chr10:10155070-10159006 12.6289
chr10:101585804-101589027 6.7638
chr10:101604155-101607416 11.1605
chr10:101619696-101622902 9.8464
chr10:10165318-10168334 10.9878
chr10:101761979-101764918 10.6036
chr10:102106502-102110676 11.3825
chr10:102151994-102156051 9.9737
chr10:102157862-102165241 9.9693
chr10:102238791-102243849 11.5089
chr10:1022613-1026620 10.2006
chr10:102360438-102363106 9.4614

Total number of rows: 18242

Table truncated, full table size 562 Kbytes.




Supplementary file Size Download File type/resource
GSM743646_382128A10_PMT525_532_grid.pair.gz 2.6 Mb (ftp)(http) PAIR
Processed data included within Sample table

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