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Status |
Public on Feb 03, 2014 |
Title |
MN clebuterol replicate 3 vs WT clebuterol replicate 1 |
Sample type |
RNA |
|
|
Channel 1 |
Source name |
MN.clenbuterol
|
Organism |
Mus musculus |
Characteristics |
strain/background: C57BL6/J genotype: myostatin null age: 7 weeks gender: male tissue: skeletal muscle (tibialis anterior) treatment: clenbuterol
|
Treatment protocol |
At 5 weeks of age, mice were given free access to either plain tap water (control) or tap water with 20 ppm clenbuterol hydrochloride (clenbuterol). Treatment duration was 2 weeks. Animals were sacrificed and tibialis anterior muscles were dissected from the right hind limbs.
|
Growth protocol |
Wild type (WT) and myostatin null (MN) mice were weaned from dams at 3 weeks of age and individually housed until 7 week of age. They were given free access to a standard chow diet.
|
Extracted molecule |
total RNA |
Extraction protocol |
Muscles were disrupted in Trizol with a TissueLyser and isolation of total RNA were carried out according to the manufacturers’ directions (Invitrogen). Isolated RNA was quantified by spectrophotometry using a spectrophotometer. Prior to microarray, RNA quality was determined using a 2100 Bioanalyzer (Agilent, Palo Alto, CA).
|
Label |
Alexa Fluor 647
|
Label protocol |
1 ug of total RNA was amplified using the amino-allyl MessageAmp II kit from Ambion. Amplified RNA was then labeled with Alexa Fluor 555 or 647 and cleaned in an RNeasy column.
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|
|
Channel 2 |
Source name |
WT.clenbuterol
|
Organism |
Mus musculus |
Characteristics |
strain/background: C57BL6/J genotype: wild type age: 7 weeks gender: male tissue: skeletal muscle (tibialis anterior) treatment: clenbuterol
|
Treatment protocol |
At 5 weeks of age, mice were given free access to either plain tap water (control) or tap water with 20 ppm clenbuterol hydrochloride (clenbuterol). Treatment duration was 2 weeks. Animals were sacrificed and tibialis anterior muscles were dissected from the right hind limbs.
|
Growth protocol |
Wild type (WT) and myostatin null (MN) mice were weaned from dams at 3 weeks of age and individually housed until 7 week of age. They were given free access to a standard chow diet.
|
Extracted molecule |
total RNA |
Extraction protocol |
Muscles were disrupted in Trizol with a TissueLyser and isolation of total RNA were carried out according to the manufacturers’ directions (Invitrogen). Isolated RNA was quantified by spectrophotometry using a spectrophotometer. Prior to microarray, RNA quality was determined using a 2100 Bioanalyzer (Agilent, Palo Alto, CA).
|
Label |
Alexa Fluor 555
|
Label protocol |
1 ug of total RNA was amplified using the amino-allyl MessageAmp II kit from Ambion. Amplified RNA was then labeled with Alexa Fluor 555 or 647 and cleaned in an RNeasy column.
|
|
|
|
Hybridization protocol |
Samples were suspended in SlideHyb#1 (Ambion), applied to slides on a Maui mixer and incubated O/N at 42ûC. After hybridization, the slides were washed sequentially in 1X SSC, 0.2% SDS; 0.1X SSC, 0.2% SDS; and 0.1X SSC for 5 minutes each, and spun dry.
|
Scan protocol |
Scanned on an Axon GenePix 4000B scanner. Images were quantified using Axon GenePix 6.0.
|
Description |
C1 Myostatin null with clenbuterol treatment (replicate 3) versus wild-type with clenbuterol treatment (replicate 1).
|
Data processing |
Median foreground intensites (no background subtraction) were normalized within arrays by print-tip loess normalization and between arrays using the scale method in limma. Spots with -100 flags were weighted zero before normalizations.
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|
|
Submission date |
Feb 28, 2011 |
Last update date |
Feb 03, 2014 |
Contact name |
Anna C Dilger |
E-mail(s) |
adilger2@illinois.edu
|
Phone |
217-333-3986
|
Fax |
217-244-5142
|
Organization name |
University of Illinois
|
Department |
Animal Science
|
Street address |
Meat Science Lab 1503 S Maryland Drive
|
City |
Urbana |
State/province |
IL |
ZIP/Postal code |
61801 |
Country |
USA |
|
|
Platform ID |
GPL9002 |
Series (1) |
GSE27577 |
Tibialis anterior muscle of myostatin null mice treated with clenbuterol |
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