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Status |
Public on Aug 18, 2011 |
Title |
HFD-L-1 |
Sample type |
RNA |
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Source name |
Mus musculus liver tissue
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Organism |
Mus musculus |
Characteristics |
strain: C57BL/6J, Gender: Male, Tissue: Liver, Age: 82weeks
|
Treatment protocol |
Starting at 56 weeks of age, fed a high-fat diet plus 30mg/kg SRT1720 (HFD-L).
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Extracted molecule |
total RNA |
Extraction protocol |
RNA was extracted using Qiagen RNAeasy Mini kit
|
Label |
Streptavidin-Cy3 bound to biotin-labeled cRNA.
|
Label protocol |
standard Illumina protocol using Illumina TotalPrep RNA Amplification Kit (Ambion; Austin, TX, cat # IL1791) In short, 0.5g of total RNA was first converted into single-stranded cDNA with reverse transcriptase using an oligo-dT primer containing the T7 RNA polymerase promoter site and then copied to produce double-stranded cDNA molecules. The double stranded cDNA was cleaned and concentrated with the supplied columns and used in an overnight in-vitro transcription reaction where single-stranded RNA (cRNA) was generated and labeled by incorporation of biotin-16-UTP.
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Hybridization protocol |
standard Illumina protocol. In short, a total of 0.75?g of biotin-labeled cRNA was hybridized at 58 degrees C for 16 hours to Illumina's Sentrix MouseRef-8 Expression BeadChips (Illumina, San Diego, CA). Each BeadChip has 24,000 well-annotated RefSeq transcripts with approximately 30-fold redundancy. The arrays were washed, blocked and the labeled cRNA was detected by staining with streptavidin-Cy3.
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Scan protocol |
Arrays were scanned at a resolution of 0.8um using the Beadstation 500 X from Illumina.
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Description |
n/a
|
Data processing |
Data was extracted using the Illumina BeadStudio software(v1). Any spots at or below the background were filtered out using an Illumina detection p-value score of 0.02 and above. The natural log of all remaining genes were used to find the avg and std of each array and the z-score normalization was calculated and presented below. Z-score = (raw value - avg)/std. Complete data including detection scores will be included in the supplemental files.
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|
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Submission date |
Nov 19, 2009 |
Last update date |
Jun 22, 2020 |
Contact name |
Supriyo De |
Organization name |
NIA-IRP, NIH
|
Department |
Laboratory of Genetics and Genomics
|
Lab |
Computational Biology & Genomics Core
|
Street address |
251 Bayview Blvd
|
City |
Baltimore |
State/province |
Maryland |
ZIP/Postal code |
21224 |
Country |
USA |
|
|
Platform ID |
GPL6885 |
Series (1) |
GSE19102 |
SRT1720 extends healthspan and lifespan in diet-induced obese mice |
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