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Sample GSM624443 Query DataSets for GSM624443
Status Public on Oct 10, 2012
Title TC01 bile_1
Sample type RNA
Source name Cultured cells
Organism Lacticaseibacillus casei
Characteristics agent: bile
strain: TC01. L. casei BL23 with a complete deletion of gene LCABL_02080.
Treatment protocol Cells were grown in MRS medium at 37C without shaking until O.D. 595 nm. reached 0.5. The culture was then two fold diluted with prewarmed MRS medium supplemented with 0.2% bovine bile (Sigma) and incubation continued for 45 min.
Growth protocol Cells were stored at -80C in MRS medium supplemented with 15% glycerol (stock culture). For the assay, cells from the stock culture were inoculated in MRS agar plates. A single colony was used to inoculate 5 ml of MRS medium and the culture was incubated overnight at 37C. This culture was used to inoculate MRS medium at 0.05 O.D. 595 nm. for the treatment or control assays.
Extracted molecule total RNA
Extraction protocol Total RNA was isolated from L. casei by using TRIZOL reagent (Invitrogen). Cells were collected by centrifugation, washed with cold 50 mM EDTA (pH 8.0), resuspended in 1 ml of TRIZOL reagent, and then mechanically disrupted with glass beads in a cell disrupter (Savant Instruments, Holbrook, N.Y.). Additional steps were performed according to the instructions of the TRIZOL manufacturer. RNA samples purified as described above were treated with the Ambion Turbo DNA-free kit (Applied Biosystems) using the routine DNase I treatment outlined by the supplier in order to remove contaminating DNA. The quality and concentration of the RNA samples were subsequently evaluated by using the Experion automated electrophoresis system (Bio-Rad). Samples with 23S/16S ratios lower than 0.85 were discarded.
Label 33P-dCTP
Label protocol First-strand cDNA was synthesized from 45 µg RNA using the Transcriptor first strand cdna synthesis kit (Roche) with random hexamer primers as recommended by the manufacturer. Labelled cDNA was purified with illustra MicroSpin G-50 Columns (GE Healthcare) as recommended by the manufacturer.
Hybridization protocol SDS based hybridization solution, during 24 hours
Scan protocol FUJIFILM FLA-3000
Data processing The software for image analysis is ArrayVision 7.0 (Imaging Research Inc.). This software yields raw and background substracted data.
The Background Substracted Signals are raw individual data. After considering the three replicates performed in the experiment and after median normalization, first between replicates and after between conditions, we obtain the ratio values for each gene/ORF (see supplementary file on Series record).
Submission date Nov 17, 2010
Last update date Oct 10, 2012
Contact name Manuel Zúñiga
Phone 34 963900022
Fax 34 963636301
Organization name Instituto de Agroquímica y Tecnología de Alimentos
Department Biotecnología
Lab Bacterias lácticas y Probióticos
Street address Avda. Profesor Agustín Escardino 7
City Paterna
State/province Valencia
ZIP/Postal code 46980
Country Spain
Platform ID GPL11074
Series (1)
GSE25448 Comparison of the response to bile of Lactobacillus casei BL23 and its derivative strain TC01

Data table header descriptions
TC01 bile-1 Raw signal for L. casei TC01 strain grown in MRS medium supplemented with bile replicate 1
VALUE Background substracted signal

Data table
ID_REF TC01 bile-1 VALUE
R1 - C1 : 1 64.14 54.008
R1 - C1 : 2 71.01 60.88
R1 - C1 : 3 25.98 15.855
R1 - C2 : 1 80.53 71.311
R1 - C2 : 2 23.65 14.43
R1 - C2 : 3 32.79 23.569
R1 - C3 : 1 82.38 72.682
R1 - C3 : 2 91.3 81.604
R1 - C3 : 3 35.16 25.461
R1 - C4 : 1 83.24 74.616
R1 - C4 : 2 37.03 28.401
R1 - C4 : 3 48.39 39.762
R1 - C5 : 1 61.1 52.918
R1 - C5 : 2 15.25 7.07
R1 - C5 : 3 8.19 0.003
R1 - C6 : 1 57.49 48.967
R1 - C6 : 2 16.01 7.482
R1 - C6 : 3 65.56 57.041
R1 - C7 : 1 177.69 168.957
R1 - C7 : 2 22.89 14.161

Total number of rows: 2688

Table truncated, full table size 66 Kbytes.

Supplementary data files not provided
Processed data are available on Series record

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