Primerscript rt pcr kit
The PrimerScript RT-PCR kit is a reagent kit designed for reverse transcription and polymerase chain reaction (RT-PCR) analysis. The kit provides the essential components for performing sensitive and accurate RNA detection and quantification.
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27 protocols using primerscript rt pcr kit
RNA Extraction, Reverse Transcription, and qPCR Analysis
Quantification of miR-486 and Its Target Genes
The expression of miR-486 and the 5S internal control gene were quantified using real-time PCR quantification and the Hairpin-it miRNAs qPCR Quantitation Kit (GenePharma, Shanghai, China) according to the manufacturer’s protocol. Specific primers for miR-486 and 5S were designed by GenePharma. The expression of these genes was analyzed using a 7300 Fast Real-Time PCR System (Applied Biosystems, Foster City, CA, USA).
The expression of PTEN, AKT, mTOR, and β-actin was quantified using Real-Time-PCR quantification and SYBR Premix Ex Taq II (TaKaRa) according to the manufacturer’s instructions. β-actin was used as a loading control. The primers that were used for qRT-PCR are listed in
Quantitative Analysis of TopBP1 Expression
Quantifying miRNA and mRNA Levels
miRNA Expression Analysis via qPCR
RNA Isolation and RT-qPCR Analysis
Gene Expression Analysis by qPCR and Western Blot
As for Western blotting, the cells were lysed in RIPA lysis buffer (CWBIO, Beijing, China) with protease and phosphatase inhibitors (CWBIO). After identifying quantities of proteins using Pierce BCA Protein Assay Kit (Invitrogen), the proteins were electrophoresed using SDS‐PAGE and transferred to PVDF membranes (Merck, Burlington, Massachusetts, USA), blocked and incubated with primary antibodies at 4°C overnight. Primary antibodies specific to HSF1, LEF1, MMP9, E2F2, E‐cadherin, N‐cadherin, vimentin, PRMT5, FLAG, GAPDH are listed in Supplementary Table
Quantitative RT-PCR Analysis of Gene Expression
Intestinal Total RNA Extraction and qRT-PCR
Quantitative Assessment of RNA Levels
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