qPCR was conducted using 12 new samples of EAT (n = 6 CAD and n = 6 non-CAD) to validate select DEmiRNAs (hsa-146b-5p, hsa-26b-5p, hsa-21-5p, hsa-320a) in the EAT of patients with CAD. miRNAs were isolated from the EAT of all patients using a commercially available miRNA isolation kit (QIAGEN, Germantown, MD, USA). Reverse transcription of the miRNAs was conducted using the miRCURY LNA RT Kit (QIAGEN, Germantown, MD, USA). PCR primers for hsa-miR-146b-5p, hsa-miR-26b-5P, hsa-miR-21-5p, and hsa-miR-320a were obtained from Qiagen. qPCR was conducted on a CFX Connect Real-Time System (Bio-Rad, Hercules, CA, USA) using the cycle settings recommended in the miRCURY LNA RT Kit (QIAGEN, Germantown, MD, USA). ΔCt values were determined for each miRNA in each sample using the spliceosomal RNA U6 for normalization. Unpaired t-test (using ΔCt values) was used to determine statistically significant differential expression of the miRNAs between patients with and without CAD.
Hsa mir 26b 5p
Manufactured by Qiagen
Hsa-miR-26b-5P is a microRNA (miRNA) product offered by Qiagen. miRNAs are small, non-coding RNA molecules that play a role in the regulation of gene expression. The core function of Hsa-miR-26b-5P is to detect and quantify the expression levels of the corresponding miRNA in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Mirna isolation kit, by Qiagen (1 mentions)
Cfx connect real time system, by Bio-Rad (1 mentions)
Mircury lna rt kit, by Qiagen (1 mentions)
Hsa mir 126 3p, by Qiagen (1 mentions)
Snord68, by Qiagen (1 mentions)
Mircury lnatm rt kit, by Qiagen (1 mentions)
Cfx96 real time pcr analysis system, by Bio-Rad (1 mentions)
Mircury lnatm sybr green pcr kit, by Qiagen (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
Miscript sybr green pcr kit, by Qiagen (1 mentions)
Miscript 2 rt kit, by Qiagen (1 mentions)
2 protocols using hsa mir 26b 5p
1
Validating DEmiRNAs in Coronary Artery Disease
2
Quantitative Analysis of miRNA Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Total RNA was extracted from tissues or cell samples using TRIzol (Invitrogen).
Complementary DNA (cDNA) was synthesized from the total RNA using miRCURY LNATMRT kit (Qiagen). qRT-PCR was performed using the miRCURY LNATM SYBR Green PCR
kit (Qiagen) and a CFX96 real-time PCR analysis system (Bio-Rad). LNATM PCR
primer mix, hsa-miR-26b-5p (GeneGlobe ID: YP00204172), hsa-miR-126-3p (GeneGlobe ID:
YP00204227), and hsa-miR-320a (GeneGlobe ID: YP00206042), and hsa-mir-744 (GeneGlobe ID:
YP00204663) were purchased from Qiagen. The miRNA expression was normalized to that of the
U6 small nuclear RNA (Qiagen, #YP00203907). Plasma miRNA expression was normalized to the
spike-in control (Qiagen).
For precursor miRNA analysis, cDNA was synthesized from total RNA with a miScript II RT
Kit (Qiagen). The expression levels of precursor miRNA were determined using a miScript
SYBR Green PCR Kit (Qiagen). cDNA was used as a template for quantitative real-time PCR
with an Hs-mir-26b-1-PR miScript Precursor Assay (#MP00001694), Hs-mir-126-1-PR miScript
Precursor Assay (#MP00000476), and Hs-mir-320a-1-PR-miScript Precursor Assay (#MP00001869;
Qiagen). Pre-miRNA expression was normalized to the control RNA (SNORD68, Qiagen;
GeneGlobe ID: MS00033712).
Complementary DNA (cDNA) was synthesized from the total RNA using miRCURY LNATMRT kit (Qiagen). qRT-PCR was performed using the miRCURY LNATM SYBR Green PCR
kit (Qiagen) and a CFX96 real-time PCR analysis system (Bio-Rad). LNATM PCR
primer mix, hsa-miR-26b-5p (GeneGlobe ID: YP00204172), hsa-miR-126-3p (GeneGlobe ID:
YP00204227), and hsa-miR-320a (GeneGlobe ID: YP00206042), and hsa-mir-744 (GeneGlobe ID:
YP00204663) were purchased from Qiagen. The miRNA expression was normalized to that of the
U6 small nuclear RNA (Qiagen, #YP00203907). Plasma miRNA expression was normalized to the
spike-in control (Qiagen).
For precursor miRNA analysis, cDNA was synthesized from total RNA with a miScript II RT
Kit (Qiagen). The expression levels of precursor miRNA were determined using a miScript
SYBR Green PCR Kit (Qiagen). cDNA was used as a template for quantitative real-time PCR
with an Hs-mir-26b-1-PR miScript Precursor Assay (#MP00001694), Hs-mir-126-1-PR miScript
Precursor Assay (#MP00000476), and Hs-mir-320a-1-PR-miScript Precursor Assay (#MP00001869;
Qiagen). Pre-miRNA expression was normalized to the control RNA (SNORD68, Qiagen;
GeneGlobe ID: MS00033712).
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