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Sample GSM1136115 Query DataSets for GSM1136115
Status Public on Feb 03, 2014
Title Transiently transfected hEEC from biopsy I with MIMIC_30d miRNA
Sample type RNA
 
Source name primary hEEC_miR-30d mimic
Organism Homo sapiens
Characteristics cell type: primary hEEC derived from healthy human endometrial biopsies at LH+0
transfected with: miR-30d mimic for 72hr
Growth protocol When hEEC cultures reached 50% confluency cells were transiently transfected with 50nM of either miR-30d mimic or Scramble miRNA using HiPerfect as recommended by the manufacturer (Qiagen, Valencia, CA, USA). After 72 hours post-transfection, cells were subjected to RNA extraction.
Extracted molecule total RNA
Extraction protocol To ensure that miRNA fraction will be recovered we performed RNA extraction using the miRNeasy Kit (Qiagen, Valencia, CA, USA). RNA extracted was quantified using a NanoDrop (Thermo Fisher Scientific Inc, MA, USA) spectrophotometer and the quality of RNA samples was assesed using a Nano LabChip BioAnalyzer 2100 (Agilent Technologies Inc, DE, USA).
Label Cy3-CTP
Label protocol 100ng of total RNA were mixed with spike and mixed with T7 promoter primer for 10' at 65C then cooled at 4C. Next to cDNA synthesis, First strand buffer, DTT, dNTP and RNAse block mix were added and incubated for 2 hours at 40C followed by 15' at 70C. Next for cRNA synthesis, transcription buffer, DTT, NTP, T7 RNA polymerase and Cy3-CTP were added and incubated for 2 hours at 40C and cooled for 5'. Then cRNA was purified according to Qiagen RNEasy kit protocol specifications. Yield and [cRNA] stimated. Then 1.65 ug de cRNA was used to be fragmented in presence of blocking agent and fragmentation buffer for 30' at 60C and cooled. Then sample was mixed with hybridization buffer.
 
Hybridization protocol 1.65 ug of cRNA was used to be fragmented in presence of blocking agent and fragmentation buffer for 30' at 60C and cooled. Then sample was mixed with hybridization buffer.
Scan protocol Microarrays slides were washed and scanned in Genepix Personal 4100A Scanner and scanned in Genepix Pro Software with a spot resolution of 5um in the green channel.
Description MIMIC_30d_I
each sample was previous tested for total RNA quality using total Nano RNA Labchip in an agilent bioanalyzer and with RIN above 9.0
Data processing Median intensity value of each spot data was log2 transformed and normalized using R software and libraries from bioconductor database. Next, replicated probes were merged by mean using GEPAS.
 
Submission date May 08, 2013
Last update date Feb 03, 2014
Contact name Juan M Moreno-Moya
E-mail(s) jmmormoy@gmail.com
Organization name FIVI
Street address Catedrático Agustín Escardino
City Valencia
ZIP/Postal code 46980
Country Spain
 
Platform ID GPL10332
Series (1)
GSE46721 Transcriptomic effects of miR-30d transient over-expression in human endometrial epithelial cells.

Data table header descriptions
ID_REF
VALUE Normalized signal intensity

Data table
ID_REF VALUE
1 15.99997799
2 15.99997799
3 6.948367232
4 6.753216749
5 6.665335917
6 6.434628228
7 6.375039431
8 6.375039431
9 6.355351096
10 6.257387843
11 6.312882955
12 6.415741768
13 6.375039431
14 6.491853096
15 6.394462695
16 6.33650656
17 6.554588852
18 6.52160044
19 6.721099189
20 15.63174547

Total number of rows: 45220

Table truncated, full table size 777 Kbytes.




Supplementary file Size Download File type/resource
GSM1136115_MIMIC_30d_I.gpr.gz 4.2 Mb (ftp)(http) GPR
Processed data included within Sample table

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