Agilent scanner g2505b

The microarray hybridization and analysis were performed by Kangcheng Bio-tech, Shanghai PR China. Briefly, RNA was purified from total RNA after removal of rRNA (mRNA-ONLY Eukaryotic mRNA Isolation Kit, Epicentre). Then, each sample was amplified and transcribed into fluorescent cRNA along the entire length of the transcripts without 39 bias utilizing a random priming method. The labeled cRNAs were hybridized to the Mouse LncRNA Array v3.0 (8×60 K, Arraystar). About 35,923 lncRNAs and 24,881 coding transcripts which were collected from the most authoritative databases such as RefSeq, UCSC Knowngenes, Ensembl, and many related landmark publications can be detected. The arrays were scanned by the Agilent Scanner G2505B, and the acquired array images were analyzed by Agilent Feature Extraction software (version 11.0.1.1). Quantile normalization and subsequent data processing were performed by using the GeneSpring GX v12.1 software package (Agilent Technologies, Santa Clara, CA, USA). Normalized data were log2-transformed and used for comparisons. LncRNAs and mRNAs, that is, at least 3 out of 9 samples have flags in Present or Marginal (“All Targets Value”) were chosen for further data analysis. DE lncRNAs and mRNAs with statistical significance between the two groups were identified through P-value and fold change filtering.

Sample preparation and microarray hybridization were performed by Kangcheng Bio-tech (Aksomics Inc., Shanghai, China). Briefly, RNA was purified from 1 µg total RNA following removal of rRNA (mRNA-ONLY Eukaryotic mRNA Isolation kit; Epicentre; Illumina, Inc., San Diego, CA, USA). Then, each sample was amplified and transcribed into fluorescent cRNA along the entire length of the transcripts without 39 bias utilizing a random priming method. The labeled cRNAs were hybridized onto the Human LncRNA Array (version, 2.0; 8660 K; ArrayStar, Inc., Rockville, MD, USA). Following the washing of the slides, the arrays were scanned by the Agilent Scanner G2505B (Agilent Technologies, Inc., Santa Clara, CA, USA). Agilent Feature Extraction software (version, 10.7.3.1; Agilent Technologies, Inc.) was utilized to analyze acquired array images. Quantile normalization and subsequent data processing were carried out using the GeneSpring GX software package (version, 11.5.1; Agilent Technologies, Inc.). Differentially expressed LncRNAs and mRNAs were identified through fold change filtering (fold change ≥3.0 or ≤0.5), standard student t-test (P<0.05) and multiple hypothesis testing (FDR<0.05). P-values and FDR were calculated by Microsoft Excel (Microsoft Corporation, Redmond, WA, USA) and MATLAB 8.2 (The MathWorks, Inc., Natick, MA, USA), respectively.

Gastrocnemius muscles from three 6‐month‐old and three 24‐month‐old mice were collected. Sample preparation and microarray hybridization were performed by Kangchen Bio‐tech, Shanghai P.R. China.19 Briefly, RNA was purified from 1 μg total RNA after removal of rRNA (mRNA‐ONLY Eukaryotic mRNA Isolation Kit, Epicentre). Then, each sample was amplified and transcribed into fluorescent cRNA along the entire length of the transcripts without 3′ bias utilizing a random priming method. The labelled cRNAs were hybridized onto the Mice LncRNA Array v2.0 (8 × 60 K, Arraystar, Rockville, MD, USA). After having washed the slides, the arrays were scanned by the Agilent Scanner G2505B (Agilent Technologies, Santa Clara, CA, USA). Agilent Feature Extraction software (version 10.7.3.1) was utilized to analyse acquired array images. Quantile normalization and subsequent data processing were carried out using the GeneSpring GX v11.5.1 software package (Agilent Technologies, Santa Clara, CA, USA). Differentially expressed lncRNAs was identified through fold change filtering (Fold Change ≥2.0 or ≤0.5), paired t‐test (P < 0.05) and multiple hypothesis testing (FDR < 0.05). P values and FDR were calculated by Microsoft Excel and MATLAB, respectively.