miRNA analysis was performed as described previously [27] . Total RNA was isolated from draining LNs from vehicle-treated and AEA-treated mice by using the miRNAeasy total RNA isolation kit and analyzed for purity using A260/280 readings. RNA purity for each sample was 1.7–1.9. Samples were analyzed using Affymetrix miRNA 1.0, allowing for analysis of 609 miRNA species. Target prediction was performed on microRNA.org MiRanda and MirSvR databases and pathway analysis was performed using Ingenuity Pathway Analysis [IPA] Software. miRNA differences were validated using QuantiTech Sybr Green PCR kit and miRNA primer assays [Qiagen].
Mirna primer assays
Manufactured by Qiagen
Sourced in Germany
The MiRNA primer assays are a set of reagents designed for the detection and quantification of microRNA (miRNA) expression levels. These assays provide a standardized and reliable method for analyzing miRNA profiles in biological samples. The core function of the MiRNA primer assays is to enable the amplification and measurement of specific miRNA targets through real-time PCR technology.
Automatically generated - may contain errors
3 protocols using mirna primer assays
1
Profiling miRNA in Draining Lymph Nodes
2
Quantification of miRNA and mRNA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was reverse-transcribed with the miScript Kit (QIAGEN). Expression levels of mature miRNAs and of mRNAs were measured using the Quantitect SYBR Green PCR Kit (QIAGEN). Relative quantities of miRNA expression were normalized to the gene encoding ribosomal protein S7 (RPS7). The miRNA primers were obtained as miRNA Primer Assays (QIAGEN).
3
Quantitative Assessment of miR-26b Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
miRNAs from tissues and serum samples were purified using the mirVANA miRNA isolation kit (Applied Biosystems, Foster City, CA) according to the manufacturer's instruction. The expression patterns of miR-26b tested and a housekeeping gene U6B were quantitatively assayed using reverse transcription and qRT-PCR. Stem loop cDNAs were synthesized using looped reverse transcription primer specific for each miRNA. All the materials were purchased from Exiqon. All samples were analyzed twice to confirm reproducibility. Real-time PCRs were conducted on an ABI Prism 7700 apparatus (Applied Biosystems). The expression of miR-26b was normalized to U6B RNA internal control. The relative level of miRNA expression was calculated using the ΔΔCt method.
Primers were purchased from Metabion GmbH (Planegg, Germany), and miRNA Primer assays were obtained from (Qiagen). Their sequences are presented in the Table, Supplemental Digital Content 2,http://links.lww.com/IBD/A907 .
Primers were purchased from Metabion GmbH (Planegg, Germany), and miRNA Primer assays were obtained from (Qiagen). Their sequences are presented in the Table, Supplemental Digital Content 2,
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!