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Sample GSM111125 Query DataSets for GSM111125
Status Public on Mar 01, 2008
Title Pancreas_P1_WT_rep3
Sample type RNA
 
Source name pooled pancreata, post-natal day 1, wild type
Organism Mus musculus
Characteristics Strain: B6D2
Developmental stage: post-natal day 1
Tissue: pooled pancreata
Genotype: wild type
Treatment protocol HNF6 expression was driven specifically to pancreatic endocrine cells beginning at embryonic day (e) 11.5 using a 1 kb enhancer from the 5’ pdx1 promoter region. This transgenic line, termed pdx1PBHnf6, was maintained on an inbred hybrid (B6D2) background. All mice were kept on a 12 hour light/dark cycle, fed Mouse diet 5015 (LabDiet), and given water ad lib. For embryonic analyses, the morning of the vaginal plug was considered e0.5. All mouse studies were performed in accordance with the Vanderbilt Institutional Animal Care and Use Committee guidelines under the supervision of the Division of Animal Care.
Extracted molecule total RNA
Extraction protocol Pancreata were dissected at e18.5 and postnatal day 1 in ice cold PBS and immediately placed into RNAlater (Ambion). Total pancreatic RNA was isolated using RNAqueous RNA Isolation Kit (Ambion) according to the manufacturer’s instructions. Agilent’s Bioanalyzer microfluidic assay (Agilent Technologies, Palo Alto CA) was used to assay RNA integrity. Spectrophotometric and fluorometric methods were combined to quantitate protein and nucleic acids present in the sample, and to ensure quality control of each sample. RNA samples were bioanalyzed individually and highly pure samples were pooled according to their genotype (3-5 animals per pool) in order to obtain an adequate quantity of RNA.
Label biotin
Label protocol Following quality control, the RNA was prepared for microarray analysis using the standard Affymetrix protocol (Affymetrix Inc, Santa Clara, CA). Briefly, a total of 5 µg of total RNA was reverse transcribed to double-stranded (ds) cDNA using an oligo-dT primer coupled to a T7 promoter. In vitro transcription from the ds cDNA was then carried out using T7 polymerase and incorporating biotin-modified CTP and UTP ribonucleotides.
 
Hybridization protocol The biotinylated cRNA (15 µg) was fragmented and hybridized to an Affymetrix GeneChip Mouse Genome 430 2.0 Array containing 45,000 sets of 11 to 25-mer oligomers, representing 39,000 mouse transcripts (34,000 are annotated as well-defined genes). Hybridized cRNA was detected using streptavidin coupled to phycoerythrin.
Scan protocol GeneChips were scanned using GeneChip Scanner 3000 7G Plus 2 and GeneChip Operating System (GCOS, Affymetrix, Santa Clara, CA). Default values were used to grid images (.DAT) and generate .CEL and .CHP files.
Description No additional description necessary
Data processing CEL files (raw Affymetrix data) were imported in GeneSpring 7.0 (Agilent Technologies) and transformed by RMA (Robust Multichip Analysis). The "Value" column shows this normalized signal intensity value.
 
Submission date May 25, 2006
Last update date Aug 28, 2018
Contact name Laura Crawford
Organization name Vanderbilt University
Department Diabetes, Endocrinology, & Metab Division
Lab Gannon Lab
Street address 715 PRB 2220 Pierce Ave
City Nashville
State/province TN
ZIP/Postal code 37232
Country USA
 
Platform ID GPL1261
Series (1)
GSE4926 Gene expression profiling of a mouse model of islet dysmorphogenesis
Relations
Reanalyzed by GSE119085

Data table header descriptions
ID_REF
VALUE RMA-transformed signal intensity value

Data table
ID_REF VALUE
1415670_at 1106.3577
1415671_at 1373.5302
1415672_at 2278.547
1415673_at 171.57144
1415674_a_at 486.2232
1415675_at 637.73987
1415676_a_at 2340.3687
1415677_at 500.29907
1415678_at 1217.7201
1415679_at 2485.9353
1415680_at 636.17676
1415681_at 634.97925
1415682_at 468.37122
1415683_at 1198.0728
1415684_at 336.5101
1415685_at 249.83522
1415686_at 538.70715
1415687_a_at 2526.6438
1415688_at 1529.7477
1415689_s_at 610.16565

Total number of rows: 45101

Table truncated, full table size 927 Kbytes.




Supplementary file Size Download File type/resource
GSM111125.CEL.gz 6.3 Mb (ftp)(http) CEL
GSM111125.CHP.gz 5.2 Mb (ftp)(http) CHP
Raw data provided as supplementary file
Processed data provided as supplementary file

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