Log In Sign up
The largest database of trusted experimental protocols

Miscript preamp pcr kit

Manufactured by Qiagen
Visit Supplier
Sourced in Germany, United States

The MiScript PreAMP PCR Kit is a laboratory equipment product designed for pre-amplification of microRNA (miRNA) samples prior to quantitative real-time PCR (qRT-PCR) analysis. The kit enables efficient amplification of miRNA targets from limited sample input.

Automatically generated - may contain errors

Lab products found in correlation

Miscript 2 rt kit, by Qiagen (16 mentions) Miscript sybr green pcr kit, by Qiagen (8 mentions) Mirneasy serum plasma kit, by Qiagen (7 mentions) Exorneasy serum plasma midi kit, by Qiagen (2 mentions) Exogenous cel mir 39 3p, by Qiagen (2 mentions) Mirneasy serum plasma spike in control, by Qiagen (2 mentions) Mirneasy kit, by Qiagen (2 mentions) Geneamp pcr system 2700, by Thermo Fisher Scientific (2 mentions) Abi viia 7, by Thermo Fisher Scientific (1 mentions) Rt pcr rotor gene q, by Qiagen (1 mentions) Mx3000p qpcr system, by Agilent Technologies (1 mentions) Hispec buffer, by Qiagen (1 mentions) Miscript hispec buffer, by Qiagen (1 mentions) Sybr green master mix, by Thermo Fisher Scientific (1 mentions) Ingenuity pathway analysis software, by Qiagen (1 mentions) 7500 real time pcr system, by Thermo Fisher Scientific (1 mentions) Bca assay, by Thermo Fisher Scientific (1 mentions) Fragment analyzer, by Agilent Technologies (1 mentions) Hiseq 4000 platform, by Illumina (1 mentions) Rneasy mini kit, by Qiagen (1 mentions)

Spelling variants of this product

MiScript PCR Kit (19 citations) MiScript PreAMP kit (3 citations) PreAMP PCR kit (2 citations)

31 protocols using miscript preamp pcr kit

1

Phenol-free RNA Extraction and miRNA Amplification

Check if the same lab product or an alternative is used in the 5 most similar protocols
RNA was isolated following a phenol-free protocol. Briefly, proteins were first precipitated using a metal cation, and then, RNA present in the supernatant was bound to an RNeasy column and washed with diluted buffers RWT and RPE and subsequently with ethanol prior to drying and elution with RNAse-free water. cDNA was pre-amplified for 12 cycles to guaranty the detection of circulating miRNAs by using the miScript PreAMP PCR Kit (Qiagen®).
Pezuk J.A., Miller T.L., Bevilacqua J.L., de Barros A.C., de Andrade F.E., e Macedo L.F., Aguilar V., Claro A.N., Camargo A.A., Galante P.A, & Reis L.F. (2017). Measuring plasma levels of three microRNAs can improve the accuracy for identification of malignant breast lesions in women with BI-RADS 4 mammography. Oncotarget, 8(48), 83940-83948.
+ Open protocol
+ Expand
2

Plasma miRNA Extraction and Amplification

Check if the same lab product or an alternative is used in the 5 most similar protocols
Plasma was obtained from blood in EDTA tubes by centrifugation at 1200 x g for 10 minutes at room temperature, and stored at -80°0°C until further use. MiRNAs were isolated from plasma using the miRNeasy Serum/Plasma Kit (Qiagen, Germany), according to the manufacturer’s protocol. Pre-amplification was performed using the miScript PreAMP PCR Kit (Qiagen) to obtain sufficient amounts of cDNA for miRNA analysis with real-time (RT) PCR.
Meeuwsen J.A., van ´t Hof F.N., van Rheenen W., Rinkel G.J., Veldink J.H, & Ruigrok Y.M. (2017). Circulating microRNAs in patients with intracranial aneurysms. PLoS ONE, 12(5), e0176558.
+ Open protocol
+ Expand
3

Profiling Prostate Cancer miRNAs in EVs

Check if the same lab product or an alternative is used in the 5 most similar protocols
A miScript II RT kit (Qiagen) was used to perform reverse transcription. The preamplification of miRNAs prior to quantification was required to accurately assess their expression because of their low abundance in EVs. A miScript Preamp PCR kit (Qiagen) in conjunction with the miScript Human Prostate Cancer PreAMP PCR primer mix was used to perform multiplex PCR-based preamplification reactions. miScript miRNA PCR array with a Human Prostate Cancer Panel (Qiagen) was used to measure miRNA expression profiles and qPCR reactions were performed on the ABI ViiA 7 (Applied Biosystems). Six control small nuclear RNAs: SNORD61, SNORD68, SNORD72, SNORD95, SNORD96A, and RNU6B/RNU6-2 were used to normalize the expression values of miRNAs, and differential expression was calculated using the ΔΔCt method. Raw data were analyzed using the miScript miRNA PCR Array Data Analysis Website (https://geneglobe.qiagen.com/us/analyze).
Chisholm J., Haas-Neill S., Margetts P, & Al-Nedawi K. (2022). Characterization of proteins, mRNAs, and miRNAs of circulating extracellular vesicles from prostate cancer patients compared to healthy subjects. Frontiers in Oncology, 12, 895555.
+ Open protocol
+ Expand
4

Serum miRNA Expression Analysis in CSU

Check if the same lab product or an alternative is used in the 5 most similar protocols
Total RNA was isolated from serum samples (200 μL per sample) of 50 CSU patients and 50 controls using a miRNeasy Serum/Plasma Kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions. Qiagen miScript Reverse Transcription (RT) Kit II (Hilden, Germany) was used for cDNA extraction. The cDNAs were amplified using the Qiagen miScript PreAMP PCR kit (Hilden, Germany). Following the cDNA extraction, the real‐time polymerase chain reaction (RT‐PCR) stage began to analyze the expression levels of miR‐155 and miR‐221. The reaction components for RT‐PCR were prepared using the Qiagen miScript SYBR Green PCR kit. SNORD61 was used for the normalization of RT‐PCR. RT‐PCR Rotor‐Gene Q (Qiagen) was used to identify SNORD61 expression levels as a reference along with the hsa‐miR‐155‐3p and hsa‐miR‐221‐3p miRNA primers.
Karstarli Bakay O.S., Demir B., Cicek D., Erol D., Aşçı Toraman Z., Gural Y, & Maurer M. (2023). In chronic spontaneous urticaria, IgE and C‐reactive protein are linked to distinct microRNAs and interleukin‐31. Clinical and Translational Allergy, 13(8), e12290.
+ Open protocol
+ Expand
5

miRNome Profiling by qPCR Array

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
miRNome profiling was performed by using miScript miRNome PCR arrays (Qiagen, MIHS-216Z). 10 ng of miRNA were reverse transcribed by using miScript II RT-PCR according to the manufacturer’s instructions. The reverse transcribed cDNA functioned as a template for the pre-amplification. Pre-amplification of mature miRNA was performed by using miScript PreAmp PCR kit (Qiagen, 331451). 10 μl of cDNA were diluted into 40 μl of H2O. 5 μl of diluted cDNA were used as a template for the pre-amplification reactions. Three different pre-amplification reactions were set up for each sample, each one using a different set of primer mix to cover the entire miRNome (Qiagen, MBHS-3216Z). Following pre-amplification, pre-amplified miRNA was pulled in one tube and used for miRNome qPCR assay. The miRNA expression was normalized to RNU6_2 housekeeping gene and the fold change to pCDH lentiviral vector control was calculated. Significantly differentiated miRNAs were analyzed for mRNA targets using DIANA mirPath software (Vlachos et al., 2015 (link)) (http://diana.imis.athena-innovation.gr/DianaTools/index.php). The predicted miRNA binding sites were determined using miRcode transcriptome-wide miRNA target prediction tool (Jeggari et al., 2012 (link)) (http://mircode.org/index.php).
Jiang Q., Isquith J., Zipeto M.A., Diep R.H., Pham J., Santos N.D., Reynoso E., Chau J., Leu H., Lazzari E., Melese E., Ma W., Fang R., Minden M., Morris S., Ren B., Pineda G., Holm F, & Jamieson C. (2019). Hyper-editing of Cell Cycle Regulatory and Tumor Suppressor RNA Promotes Malignant Progenitor Propagation. Cancer cell, 35(1), 81-94.e7.
+ Open protocol
+ Expand
6

Optimized miRNA Profiling in Total RNA

Check if the same lab product or an alternative is used in the 5 most similar protocols
Using the miScript II RT kit (Qiagen) with HiSpec Buffer (Qiagen) to focus on mature miRNAs with oligo-dT primers, we reverse-transcribed 100 ng of total RNA. Next, the cDNA was amplified with miScript PreAMP PCR kit (Qiagen). The PCR was then performed with the miScript SYBR Green PCR kit (Qiagen) on a Mx3000P Q-PCR system (Agilent Technologies), according to the manufacturer’s instructions. The PCR miScript Primer Assays (Qiagen) of let-7f (MIMAT0000067), miR-199a-3p (MIMAT0000232), miR-1207-5p (MIMAT0005871), miR-21 (MIMAT0000076), miR-24 (MIMAT0000080), miR-29a (MIMAT0000086), miR-29b (MIMAT0000100), miR-29c (MIMAT0000681), miR-34a (MIMAT0000255), miR-451 (MIMAT0001631), and RNU6-2 were used for q-RT-PCR.
Spear R., Boytard L., Blervaque R., Chwastyniak M., Hot D., Vanhoutte J., Lamblin N., Amouyel P, & Pinet F. (2019). Let-7f: A New Potential Circulating Biomarker Identified by miRNA Profiling of Cells Isolated from Human Abdominal Aortic Aneurysm. International Journal of Molecular Sciences, 20(21), 5499.
+ Open protocol
+ Expand
7

Colorectal miRNA Quantification

Check if the same lab product or an alternative is used in the 5 most similar protocols
MiRNAs from colorectal tissues and plasma were polyadenylated and reverse transcribed using the miScript II RT kit and miScript HiSpec buffer (Qiagen) following the manufacturer’s procedure. RNAse-free water (40 μL) was added to the synthesized cDNA (10 μL), and the mixture was aliquoted into PCR tubes and stored at −20°C until analysis.
MiScript PreAMP PCR Kit (Qiagen) was used to pre-amplify the cDNA target templates following manufacturer instructions. MiR-16 was analyzed to determine the optimal dilution for real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR). Samples that generated Ct values between 10 and 24 needed no further dilution for target miRNAs using RT-qPCR. Efficiency of reverse transcription was measured using miRTC assay. Efficient reverse transcription for the miRTC primer assay was set at Ct values between 14 and 20.
Fellizar A., Refuerzo V., Ramos J.D, & Albano P.M. (2022). Expression of specific microRNAs in tissue and plasma in colorectal cancer. Journal of Pathology and Translational Medicine, 57(3), 147-157.
+ Open protocol
+ Expand
8

Quantitative RT-PCR of miR-9 Precursors

Check if the same lab product or an alternative is used in the 5 most similar protocols
RT was conducted using a miScript II RT kit (Qiagen). miR-9 precursors (pre-mir-9-1, pre-mir-9-2, pre-mir-9-3) were pre-amplified using stock primers for RT-qPCR from Qiagen. After this, a 1:20 dilution of the pre-amp product was used for normal qPCR. Standard curves were prepared from 100 fmol using stocks of 9-1, 9-2, and 9-3 oligos. Pre-amplification was carried out using Qiagen miScript Precursor assay kits for pre-mir-9-1, -9-2, and -9-3 respectively, along with a miScript PreAMP PCR Kit (Qiagen).
Mead E.A., Wang Y., Patel S., Thekkumthala A.P., Kepich R., Benn-Hirsch E., Lee V., Basaly A., Bergeson S., Siegelmann H.T, & Pietrzykowski A.Z. (2023). miR-9 utilizes precursor pathways in adaptation to alcohol in mouse striatal neurons. Advances in Drug and Alcohol Research, 3, 11323.
+ Open protocol
+ Expand
9

Exosome RNA Isolation and miRNA Profiling

Check if the same lab product or an alternative is used in the 5 most similar protocols
Total RNA, including miRNAs, was isolated from serum exosomes using the commercial exoRNeasy serum plasma midi kit (Qiagen, Germany) according to the manufacturer’s instructions. Exogenous cel-miR-39-3p (Qiagen) was added to the samples to measure the RNA isolation efficiency, as determined by the manufacturer.
The complementary DNA (cDNA) was then synthesized from 100 ng of total RNA by using the miScript® II RT kit (Qiagen), and miRNA preamplification was performed using the miScript® PreAmp PCR kit (Qiagen) according to the manufacturer’s instructions.
Da-Silva C.C., Anauate A.C., Guirao T.P., Novaes A.D., Maquigussa E, & Boim M.A. (2022). Analysis of exosome-derived microRNAs as early biomarkers of lipopolysaccharide-induced acute kidney injury in rats. Frontiers in Physiology, 13, 944864.
+ Open protocol
+ Expand
10

Quantitative miRNA expression in CSF

Check if the same lab product or an alternative is used in the 5 most similar protocols
RT was performed on 5 μL of miRNA using the miScript II RT Kit. One microliter of complementary DNA was preamplified using the miScript PreAmp PCR Kit following the manufacturer's recommendations (Qiagen). The spike-in control C. elegans miR-39 mimic was not preamplified. qPCR reactions were performed in duplicate, except for miR-24-3p in CSF, a putative internal reference gene,14 (link) which was assayed in triplicate. Patients' subgroups and controls were equally distributed across each run to minimize interrun bias for a single miRNA target.
Perdaens O., Dang H.A., D'Auria L, & van Pesch V. (2020). CSF microRNAs discriminate MS activity and share similarity to other neuroinflammatory disorders. Neurology® Neuroimmunology & Neuroinflammation, 7(2), e673.
+ Open protocol
+ Expand

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

Sign up now

Revolutionizing how scientists
search and build protocols!