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| Status |
Public on Jun 08, 2018 |
| Title |
RIF woman serum Replicate 3 |
| Sample type |
RNA |
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| Source name |
RIF woman serum Replicate 3
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| Organism |
Homo sapiens |
| Characteristics |
Sex: female
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| Treatment protocol |
After 1 h incubation, serum was separated by centrifugation, aliquoted and stored at −70 °C until further analyses.
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| Growth protocol |
10 ml of venous blood was withdrawn into anticoagulant-free tubes.
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| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using TRIzol (Invitrogen) and purified with RNeasy mini kit (QIAGEN) according to manufacturer’s instructions. RNA quality and quantity was measured by using nanodrop spectrophotometer (ND-1000, Nanodrop Technologies) and RNA Integrity was determined by gel electrophoresis. miRNA labeling and array hybridization
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| Label |
Hy3
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| Label protocol |
The miRCURY LNA Array system (Exiqon, Vedbaek, Denmark) was used according to the manufacturer’s guideline for miRNA labelling. 1, After quality control, the miRCURY™ Hy3™/Hy5™ Power labeling kit (Exiqon, Vedbaek, Denmark) was used according to the manufacturer’s guideline for miRNA labelling by following steps: a, 1μLRNA in 2.0 μL of water was combined with 1.0 μL of CIP buffer and CIP (Exiqon). The mixture was incubated for 30 min at 37°C. b, The Reaction was terminated by incubation for 5 min at 95°C. Then 3.0 μL of labeling buffer, 1.5 μL of fluorescent label (Hy3TM), 2.0 μL of DMSO, 2.0 μL of labeling enzyme were added into the mixture.The labeling reaction was incubated for 1 h at 16°C c, Terminated by incubation for 15 min at 65°C. 2, After stopping the labeling procedure, the Hy3™-labeled samples were hybridized on the miRCURYTM LNA Array (v.18.0) (Exiqon) according to array manual. a, The total 25 μL mixture from Hy3™-labeled samples with 25 μL hybridization buffer were first denatured for 2 min at 95°C, incubated on ice for 2 min b, Then hybridized to the microarray for 16–20 h at 56°C in a 12-Bay Hybridization Systems (Hybridization System - Nimblegen Systems, Inc., Madison, WI, USA) c, Following hybridization, the slides were achieved, washed several times using Wash buffer kit (Exiqon)
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| Hybridization protocol |
Each array was hybridized with 100 ng of Cy3-labelled RNA using the miRNA Complete Labeling and Hyb Kit (Exiqon) in a hybridization oven that was set at 55 ℃ and 20 r.p.m for 20 h according to the manufacturers’ instructions.
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| Scan protocol |
The slides were scanned using the Axon GenePix 4000B microarray scanner (Axon Instruments, Foster City, CA).
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| Description |
serum were obtained from women who had experienced RIF after ovarian stimulation and IVF-ET. RIF was defined as women who had undergone≥3 failed cycles with high-grade embryos were transferred.
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| Data processing |
Scanned images were then imported into GenePix Pro 6.0 software (Axon) for grid alignment and data extraction. Replicated miRNAs were averaged and miRNAs that intensities>=30 in all samples were chosen for calculating normalization factor.
Expressed data were normalized using the Median normalization. After normalization, significant differentially expressed miRNAs between two groups were identified through Fold change and P-value. Differentially expressed miRNAs between two samples were filtered through Fold change. Finally, hierarchical clustering was performed to show distinguishable miRNA expression profiling among samples.
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| Submission date |
Oct 30, 2017 |
| Last update date |
Jun 08, 2018 |
| Contact name |
lin liu |
| E-mail(s) |
linliu@mail.ustc.edu.cn
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| Phone |
86-931-8356307
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| Organization name |
lanzhou university
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| Street address |
chengguanqu donggangxilu 1 hao
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| City |
lanzhou |
| ZIP/Postal code |
0931730000 |
| Country |
China |
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| Platform ID |
GPL19128 |
| Series (1) |
| GSE106307 |
human serum: Control vs. patients with repeated implantation failure |
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| Supplementary file |
Size |
Download |
File type/resource |
| GSM2835783_6.gpr.gz |
962.0 Kb |
(ftp)(http) |
GPR |
| Processed data are available on Series record |
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