Reverse transcription of isolated miRNA from ten laryngeal squamous cell carcinoma, patient blood samples and ten controls was performed according to the manufacturer’s recommendations using Megaplex™RT Primers Human Pool A and B, TaqMan® Human MicroRNA Array A and B purchased from Applied Biosystems using a 7900 HT System (Applied Biosystems). The expression levels of 377 human miRNA genes for each group were analysed. The following miRNAs revealed an altered expression profile: miR-122, miR-21, miR-155, miR-222, miR-181a, LET-7a, miR-31, miR-141, miR-149, miR-182, miR-145, miR-223, miR-133a, miR-485, miR-122, miR-33. These were chosen for further investigations.
Taqman human microrna array a and b
Manufactured by Thermo Fisher Scientific
Sourced in United States
The TaqMan® Human MicroRNA Array A and B are high-throughput PCR arrays designed for the analysis of human microRNA expression. The arrays contain pre-designed and validated assays for the simultaneous measurement of hundreds of human microRNAs in a single reaction. The arrays provide a comprehensive and sensitive tool for investigating human microRNA profiles.
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Lab products found in correlation
Megaplex rt primers human pool a and b, by Thermo Fisher Scientific (4 mentions)
7900ht system, by Thermo Fisher Scientific (4 mentions)
Taqman microrna reverse transcription kit, by Thermo Fisher Scientific (2 mentions)
Taqman preamp kit, by Thermo Fisher Scientific (1 mentions)
Rq manager 1, by Thermo Fisher Scientific (1 mentions)
Taqman low density array human microrna panel, by CapitalBio (1 mentions)
Mirneasy serum plasma kit, by Qiagen (1 mentions)
Mirvana mirna mimic cel mir 39, by Thermo Fisher Scientific (1 mentions)
Taqman microrna assay, by Thermo Fisher Scientific (1 mentions)
7900ht real time pcr system, by Thermo Fisher Scientific (1 mentions)
7900ht fast real time pcr system, by Thermo Fisher Scientific (1 mentions)
7 protocols using taqman human microrna array a and b
1
Profiling Laryngeal Cancer miRNA
2
Profiling miRNA in Congenital Hemochromatosis and AMD
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Reverse transcription of isolated miRNA from 5 patients with congenital hemochromatosis and AMD, 5 AMD patients without hemochromatosis, and 5 healthy controls were prepared according to the manufacturer’s recommendation using the Megaplex™RT Primers Human Pool A and B, and the TaqMan® Human MicroRNA Array A and B purchased from Applied Biosystems using a 7900 HT System (Applied Biosystems). The expression levels of 377 human miRNA genes for each group were analyzed. miRNA genes that revealed an altered expression profile and were homological to the 3′-UTRs of iron-related genes according to the DIANA database, were chosen for further investigations.
3
Serum miRNA Profiling in Bullous Pemphigoid
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For screening, a qPCR-based array was performed as previously described [16 (link)]. Briefly, total RNA was isolated from serum samples of the 20 active BP patients and 20 healthy controls with a miRNeasy Serum/Plasma Kit (Qiagen, Hilden, Germany). The yield of total RNA was 7-15 ng/μL. The cDNA was obtained from 3 μL of RNA with use of the TaqMan® MicroRNA Reverse Transcription Kit (Applied Biosystems, Beverly, USA). The TaqMan® PreAmp Kit (Applied Biosystems) was used for preamplifying with 2.5 μL of cDNA product per specimen. The qPCR-based array analysis was performed with use of the TaqMan Low Density Array Human MicroRNA Panel (CapitalBio, Beijing, China). A total of 768 known miRNAs were quantified with use of the TaqMan® Human MicroRNA Array A and B (Applied Biosystems). Raw cycle threshold (Ct) values were calculated by SDS 2.4 and RQ manager 1.2 software (Applied Biosystems). A Ct value of ≥35.0 was used as a cut-off.
4
Differential miRNA Expression in HCC
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The expression of 377 miRNAs for five HCC patients and five healthy controls was analyzed using the MegaplexTMRT Primers Human Pool A and B and the TaqMan Human MicroRNA Array A and B (Applied Biosystems) on a 7900 HT System (Applied Biosystems). Twenty-eight different miRNA genes, which were expressed in the cancer patients, but not in the controls, were chosen for further investigation.
miRNA conversion to cDNA was achieved by the TaqMan MicroRNA Reverse Transcription kit (Applied Biosystems) and the obtained cDNA was stored at –80°C. During the analysis using TaqMan MicroRNA Assay (Thermo Fisher Scientific), 5 nmol mirVana miRNA Mimic (cel-miR-39) (Ambion) was used as an endogenous control. The reaction was performed as follows: 2 min at 50°C and 10 min at 95°C, then 40 cycles consisting of 95°C for 30 s, 60°C for 30 s, and 72°C for 60 s. Data were analyzed in the ABI Prism 7000 sequence detection system (SDS Software). Each assay included samples without RT and cDNA. The 2–∆∆Ct method was used for calculations of relative gene expression levels [23 (link), 24 (link)].
miRNA conversion to cDNA was achieved by the TaqMan MicroRNA Reverse Transcription kit (Applied Biosystems) and the obtained cDNA was stored at –80°C. During the analysis using TaqMan MicroRNA Assay (Thermo Fisher Scientific), 5 nmol mirVana miRNA Mimic (cel-miR-39) (Ambion) was used as an endogenous control. The reaction was performed as follows: 2 min at 50°C and 10 min at 95°C, then 40 cycles consisting of 95°C for 30 s, 60°C for 30 s, and 72°C for 60 s. Data were analyzed in the ABI Prism 7000 sequence detection system (SDS Software). Each assay included samples without RT and cDNA. The 2–∆∆Ct method was used for calculations of relative gene expression levels [23 (link), 24 (link)].
5
miRNA Expression Profiling of Cell Cycle Phases
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RNA isolated from two samples of G1 and three samples of S and G2 phase-sorted NCI-H295R cells were studied using TaqMan Low Density Array (TLDA) cards, according to the manufacturer’s instructions. The miRNA expression profiling using TLDA was performed as previously reported [55 (link)]. 30 ng of total RNA was reverse transcribed and pre-amplified using Megaplex RT primer pool A and B and Megaplex PreAmp primers, respectively. Quantitative real-time PCR were carried out in TaqMan Human MicroRNA Array A and B on a 7900HT Real time PCR System (Applied Biosystems by Life Technologies).
6
Screening miRNA expression in endometriosis
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miRNA expression was first screened using TaqMan® Human MicroRNA Array A and B (Applied Biosystems, Foster City, CA, USA). Megaplex™RT Primers Human Pool A and B were purchased from Applied Biosystems and used to prepare the reverse transcription reaction according to manufacturer’s instructions. Amplifications were performed on a 7900HT Fast Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) under the following conditions: 2 min at 50 °C, 10 min at 94.5 °C, and 40 cycles each for 30 s at 97 °C and 1 min at 57 °C. The expression levels of 754 human miRNA genes were assessed in the first step in the screening cohort of randomly chosen FFPE endometriosis samples (OE) (n = 20) and controls (CG) (n = 20) and in ovarian cancer samples (n = 20 for high-grade ovarian cancer + 20 for endometrioid ovarian cancer). miRNAs with altered expression profiles (based on 2-fold change, PCR amplification curves, and confirmed in the literature) were chosen for further investigations in the examined groups of patients: DIE, SE, OE, and CG.
7
Profiling Human miRNA Expression in AMD
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Megaplex™RT Primers Human Pool A and B were purchased from Applied Biosystems and used to prepare the reverse transcription reaction according to manufacturer’s recommendation. Real-time PCR was performed using a 7900 HT System (Applied Biosystems) using TaqMan® Human MicroRNA Array A and B purchased from Applied Biosystems. The expression levels of 377 human miRNA genes were assessed in AMD patients and controls. miRNA with altered expression profiles were chosen for further investigations.
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