|
| Status |
Public on Dec 20, 2010 |
| Title |
LD muscle_Ujumqin_100d(dpc)_rep 2 |
| Sample type |
RNA |
| |
|
| Source name |
LD muscle,Ujumqin,100 day(dpc),replicate 2
|
| Organism |
Ovis aries |
| Characteristics |
tissue: skeletal muscle
development stage: 100 day(dpc)
breed: Ujumqin
gender: female
|
| Treatment protocol |
The longissimus dorsi from every fetus were dissected and weighed.The first sample (3 g) was dissected and frozen rapidly whole in isopentane chilled over liquid nitrogen for histology and immunohistochemistry examination. The second sample (5 g) was snap frozen in liquid nitrogen for gene expression analysis.
|
| Growth protocol |
All purebred female animals involved were selected based on their age (3-5 years old), body weight (50-55kg) and body size. These selected animals were subjected to pre-feeding for 45 days. Seventy eight Ujumqin and fifty four Texel ewes were prepared for the study. All ewes were synchronized for estrus by implanted CIDR (Pharmacia & Upjohn Pty Limited, NSW, and AU) and intramuscular injection PMSG (Ningbo Renjian Pharmaceutical Co., Ltd, CN) disposal according to the manufacturer’s protocol, followed by artificial insemination using the corresponding breed sires’ sperm. Three pregnant ewes were subject to caesarean section to collect fetuses from Texel and Ujumqin sheep at five fetal development stages (70, 85, 100, 120, 135d of gestation) and eight development stages (70, 85, 100, 120, 135d of gestation, birth, 1 month and 2 month), respectively.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from the frozen longissimus dorsi using an RNeasy Mini Kit with DNase treatment (Qiagen, Valencia, CA, USA) according to the manufacturer’s protocol. The RNA quantity and quality were measured by ND-1000 (Nanodrop, Wilmington, DE, USA) and 2100 Bioanalyzer (Agilent, Santa Clara, CA,USA), respectively.
|
| Label |
Cy3
|
| Label protocol |
Target preparation was performed using a Low Input Quick Amp Labeling Kit, one-color (Agilent) according to the manufacturer's protocol. The total RNA extracted from longissimus dosi muscles from each individual were labeled with Cy3 fluorescence.
|
| |
|
| Hybridization protocol |
The generated targets were mixed and subjected to hybridization to the Sheep Gene Expression Microarray (Agilent) according to the manufacturer's protocol.
|
| Scan protocol |
Scanning of the microarrays was done by a DNA microarray scanner (Agilent). Some important scanning parameters were: 5μm of scan resolution, PMT with 100% and10% one time respectively. All probe sets whose signals were reported “absent” flag call on all 31arrays were filtered out, and those from probe sets called “present” on at least one chip were used for further analysis.
|
| Description |
Gene expression in fetal LD muscle at 100d(dpc) in Ujumqin sheep
|
| Data processing |
Scanner output image files were normalized and filtered using Feature Extraction Software v10.5 (Agilent) and the quantile method was used for data normalization.
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| |
|
| Submission date |
Aug 11, 2010 |
| Last update date |
Dec 20, 2010 |
| Contact name |
Lixin Du |
| E-mail(s) |
hxren306@sina.com
|
| Phone |
86-010-62816002
|
| Fax |
86-010-62818815
|
| Organization name |
Chinese Academy of Agricultural Sciences
|
| Department |
Institute of Animal Sciences
|
| Lab |
National Center for Molecular Genetics and Breeding of Animal
|
| Street address |
2 # Yuanmingyuan West Rd
|
| City |
Beijing |
| ZIP/Postal code |
100193 |
| Country |
China |
| |
|
| Platform ID |
GPL10778 |
| Series (1) |
| GSE23563 |
Profiles of gene expression in skeletal muscle in sheep from the middle gestation to postnatal stages |
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