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Status |
Public on Feb 09, 2017 |
Title |
Amniotic fluid_E13 mice |
Sample type |
RNA |
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Source name |
Amniotic fluid, E13
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Organism |
Mus musculus |
Characteristics |
strain/background: C57BL/6 tissue: amniotic fluid developmental stage: E13
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Treatment protocol |
After three days of acclimatization period, two females were kept with one male for 12 h during the night. In the morning of the test day, the simultaneous presence of vaginal plug and spermatozoa in vaginal cytology was considered as day 0 of gestation. The pregnancy was further confirmed by weight increase of female mice.
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Growth protocol |
All mice were raised in pathogen-free room and the room was maintained at a constant temperature of 23±1℃ with a 12 h light/dark cycle. Food and water were available ad libitum in the home cages.
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Extracted molecule |
total RNA |
Extraction protocol |
Cellular fractions of amniotic fluid was collected by centrifugation at 1000 g for 10 min and then the total RNA was harvested using TRIzol (Invitrogen, Carlsbad, CA, USA) and RNeasy mini kit (QIAGEN) according to the manufacturers' instructions.
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Label |
Hy3
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Label protocol |
The miRCURY™ Hy3™/Hy5™ Power labeling kit (Exiqon, Vedbaek, Denmark) was used according to the manufacturer’s guideline for miRNA labelling.
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Hybridization protocol |
The Hy3TM-labeled samples were hybridized on the miRCURYTM LNA microRNA Array, 6th generation (Exiqon) according to array manual.
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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 |
AF13 Experimental animals were commercially-available C57BL/6 mice (SLAC, Shanghai, China).
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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 >=50 in all samples were chosen for calculating normalization factor. Expressed data were normalized using the Median normalization. After normalization, differentially expressed miRNAs with statistical significance were identified through Volcano Plot filtering.
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Submission date |
Jun 26, 2015 |
Last update date |
Feb 09, 2017 |
Contact name |
Ting ting Sun |
E-mail(s) |
suntingting1218@126.com
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Organization name |
Zhejiang University School of Medicine
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Department |
Institute of Neuroscience and Anatomy
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Lab |
C115
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Street address |
866 Yuhangtang Road
|
City |
Hangzhou |
State/province |
Zhejiang |
ZIP/Postal code |
310058 |
Country |
China |
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Platform ID |
GPL20632 |
Series (1) |
GSE70324 |
microRNAs profiling of amniotic fluid: evidence of the synergism of microRNAs in directing axon guidance during development |
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