The microarray analysis for miRNA profiling was conducted by the miRCURY LNA Array system (Exiqon, Vedbaek, Denmark). Total RNA was extracted and purified, using the mirVana miRNA Isolation Kit (Ambion, Austin, TX, USA), following the manufacturer's instructions. Each array was hybridized with either Hy3 or Hy5-labeled RNA, using the miRNA Complete Labeling and Hyb Kit (Exiqon) in a hybridization oven. After hybridization, each array was washed and scanned and the raw data were subjected to background subtraction and normalization with the limma R-package [39 (link)]. Triplicate miRNA probes were averaged and their intensities (>=30) in all samples were saved for calculating normalization factor using the Median normalization method. Discriminant miRNAs and differences between groups were analyzed using Bayes moderated t test (limma), with Benjamini Hochberg false discovery rate (FDR) at P < 0.05, unless otherwise specified. A cut-off of 2-fold change and FDR<0.05 was applied to select up- and down-regulated miRNAs.
Mirna complete labeling and hyb kit
Manufactured by Qiagen
The MiRNA Complete Labeling and Hyb Kit is a laboratory equipment product from Qiagen. The core function of this kit is to facilitate the labeling and hybridization of microRNA (miRNA) samples for downstream analysis.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using mirna complete labeling and hyb kit
1
miRNA Profiling by Microarray Analysis
2
Microarray Analysis for miRNA Profiling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The microarray analysis for miRNA profiling was conducted by the Shanghai Kangcheng Technology using the miRCURY LNA Array system (Exiqon, Vedbaek, Denmark). Total RNA was extracted and purified using the mirVana miRNA Isolation Kit (Ambion, Austin, TX, USA) following the manufacturer's instructions. Each array was hybridised with 100 ng of Cy3-labelled RNA using the miRNA Complete Labeling and Hyb Kit (Exiqon) in a hybridisation oven that was set at 55 °C and 20 r.p.m. for 20 h according to the manufacturer's instructions. After hybridisation, each array was washed in staining dishes with the Gene Expression Wash Buffer and scanned, and the raw data were normalised using the quantile algorithm. The threshold value for significance that was used to define upregulation or downregulation of miRNAs was a fold change >2 or <0.5, respectively.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!