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Status |
Public on Oct 15, 2013 |
Title |
Extrahepatic Bileduct and gallbladder en bloc at day 14 after normal saline injection miRNA rep2 |
Sample type |
RNA |
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Source name |
Extrahepatic Bileduct and gallbladder en bloc at day 14 after normal saline injection
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Organism |
Mus musculus |
Characteristics |
tissue: Extrahepatic Bileduct and gallbladder strain: Balb/c time: 14 days treatment: Normal saline
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Treatment protocol |
Newborn mice were injected with 20 μL of a solution containing 1.5x10^6 fluorescent-forming units of Rhesus rotavirus type-A or 0.9% NaCl (normal saline) intraperitoneally within 24 hours of birth.
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Growth protocol |
Balb/c mice were purchased from Charles River Laboratories (Wilmington, MA).
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Extracted molecule |
total RNA |
Extraction protocol |
Extrahepatic bile ducts and gallbladder were microdissected en bloc at 3, 7 and 14 days after RRV or saline injection and stabilized with RNAlater (QIAGEN, CA). 3 sets of samples were generated for each time point (experimental and control groups), with each set containing 2-6 extrahepatic bileducts. Total RNA was isolated from extrahepatic bileducts of RRV- or saline-injected mice using miRNeasy Mini Kit, according to the manufacturer’s protocol (QIAGEN, CA). RNA integrity was verified using the Agilent 2100 Bioanalyser (Agilent Technologies, Inc., CA).
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Label |
FAM
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Label protocol |
Reverse transcription into first strand cDNA was performed using reagents from TaqMan® MicroRNA RT Kit and Megaplex RT Primer Pools (Applied Biosystems, CA). Quantitative real-time PCR was performed using the TaqMan® Array Rodent MicroRNA Card v2.0 (A and B) with the Universal PCR Master Mix, No AmpErase UNG using 7900 HT Fast Real-Time PCR System (Applied Biosystems).
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Hybridization protocol |
n/a
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Scan protocol |
n/a
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Description |
Control group at day 14
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Data processing |
Expression was calculated using the comparative Ct method (Schmittgen TD, Livak KJ. Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc. 2008;3: 1101-8) Normalization was done using average of 4 MammU6 probes on each plate. The deltaCt values (Ct_Target − Ct_MammU6) were imported to GeneSpring GX 11.5 platform (Agilent Technologies, CA), converted to log2-based values and used for statistical analyses. Fold Change worksheet reports each sample/average of 9 normal controls (i.e., D3_NS_1~3, D7_NS_1~3, D14_NS_1~3) ratios.
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Submission date |
Oct 15, 2012 |
Last update date |
Oct 15, 2013 |
Contact name |
Jorge A Bezerra |
E-mail(s) |
jorge.bezerra@cchmc.org
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Organization name |
Cincinnati Children's Hospital Medical Center
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Department |
Gastroenterology, Hepatology & Nutrition
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Street address |
3333 Burnet Ave.
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City |
Cincinnati |
State/province |
OH |
ZIP/Postal code |
45229 |
Country |
USA |
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Platform ID |
GPL15053 |
Series (2) |
GSE41593 |
Taqman Low Density Arrays (TLDA) based microRNAs expression profile of mouse extrahepatic bileducts and gallbladders during a mouse model of biliary atresia. |
GSE41595 |
Integrative genomics identifies candidate microRNAs for pathogenesis of experimental biliary atresia |
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