Serum expression levels of mature miRNAs, miR-223 and miR-155 were evaluated using miScript miRNA PCR primer assays and miScript SYBER green PCR kit (Qiagen, Valenica, CA) according to the manufacturer’s protocol. The housekeeping miRNA SNORD68 was used as the internal control as previously described (Shaker and Senousy 2017 (link)). Real-time PCR was performed using Rotor gene Q System (Qiagen, Valenica, CA) using the following conditions: 95 °C for 30 min, followed by 40 cycles at 94 °C for 15 s, 55 °C for 30 s, and 70 °C for 30 s. The cycle threshold (Ct) is the number of cycles required for the fluorescent signal to cross the threshold in real-time PCR. ∆Ct was calculated by subtracting the Ct values of miRNA SNORD68 from the Ct values of the target miRNAs. Fold change was calculated using 2-∆∆Ct for relative quantification.
Miscript mirna pcr primer assays
Manufactured by Qiagen
The MiScript miRNA PCR primer assays are a set of primers designed for the detection and quantification of microRNA (miRNA) expression levels using real-time PCR technology. The assays target specific miRNA sequences and can be used to measure the expression of individual miRNAs or a panel of miRNAs in a variety of sample types.
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Lab products found in correlation
3 protocols using miscript mirna pcr primer assays
1
Quantifying Serum miRNA Levels
2
Quantification of Mature miRNA Levels
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Serum expression levels of mature miRNAs, miR-223 and miR-155 were evaluated using miScript miRNA PCR primer assays and miScript SYBER green PCR kit (Qiagen, Valenica, CA) according to the manufacturer's protocol. The housekeeping miRNA SNORD68 was used as the internal control as previously described [22] . Real-time PCR was performed using Rotor gene Q System (Qiagen, Valenica, CA) using the following conditions: 95ºC for 30 min, followed by 40 cycles at 94ºC for 15 s, 55ºC for 30 s, and 70ºC for 30 s. The cycle threshold (Ct) is the number of cycles required for the fluorescent signal to cross the threshold in real-time PCR. ∆Ct was calculated by subtracting the Ct values of miRNA SNORD68 from the Ct values of the target miRNAs. Fold change was calculated using 2 -∆∆Ct for relative quantification.
3
Quantification of Serum miRNA Levels
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Serum expression levels of mature miRNAs, miR-223 and miR-155 were evaluated using miScript miRNA PCR primer assays and miScript SYBER green PCR kit (Qiagen, Valenica, CA) according to the manufacturer's protocol. The housekeeping miRNA SNORD68 was used as the internal control. Real-time PCR was performed using Rotor gene Q System (Qiagen, Valenica, CA) with the following conditions: 95ºC for 30 min, followed by 40 cycles at 94ºC for 15 s, 55ºC for 30 s, and 70ºC for 30 s. The cycle threshold (Ct) is the number of cycles required for the fluorescent signal to cross the threshold in real-time PCR. ∆Ct was calculated by subtracting the Ct values of miRNA SNORD68 from the Ct values of the target miRNAs. Fold change was calculated using 2 -∆∆Ct for relative quantification [22] .
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