Total RNA was isolated from the tissue specimens and cultured cells using an RNeasy Plus Mini Kit (Qiagen, Chatsworth, CA). Plasma RNA was isolated with a miRNeasy Serum/Plasma Kit (Qiagen, Chatsworth, CA) according to the manufacturer’s instructions. cDNA was synthesized by reverse transcription using an MMLV transcriptase (Promega) using random primers. For gene detection, real-time RT-PCR was performed on a LC480 real-time PCR detection system (Bio-Rad) using a Roche SYBR FAST Universal qPCR Kit (Roche Molecular Biochemicals). GAPDH was used as an internal control. The sequences of primers are presented in Table 1 .
Lc480 real time pcr detection system
Manufactured by Bio-Rad
Sourced in United States
The LC480 real-time PCR detection system is a laboratory instrument designed for quantitative real-time polymerase chain reaction (qRT-PCR) analysis. The system provides precise detection and quantification of nucleic acid sequences in a sample.
Automatically generated - may contain errors
Lab products found in correlation
Sybr fast universal qpcr kit, by Roche (3 mentions)
Trizol reagent, by Thermo Fisher Scientific (2 mentions)
First strand cdna synthesis kit for rt pcr, by Roche (2 mentions)
Nanodrop 2000, by Thermo Fisher Scientific (2 mentions)
M mlv transcriptase, by Promega (1 mentions)
Rneasy plus mini kit, by Qiagen (1 mentions)
Mirneasy serum plasma kit, by Qiagen (1 mentions)
3 protocols using lc480 real time pcr detection system
1
RNA Extraction and Real-Time RT-PCR Analysis
2
RNA Extraction, RT-PCR, and Gene Expression
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Total RNA was isolated from tissue specimens using Trizol reagent (Invitrogen, Life Technologies Inc., Germany) according to the manufacturer's instructions. The density and purity of RNA were measured with NanoDrop 2000 (Thermo Fisher Scientific, USA). cDNA was synthesized by reverse transcription by First Strand cDNA Synthesis Kit for RT-PCR (Roche, Switzerland) according to the manufacturer's instructions. qRT-PCR was performed on LC480 real-time PCR detection system (Bio-Rad, Hercules, CA, USA), and a Roche SYBR FAST Universal qPCR Kit (Roche, Switzerland) was used for gene detection. All primers used for qRT-PCR were designed in primerBank (https://pga.mgh.harvard.edu/primerbank/index.html) and synthesized by BGITech (Beijing, China). GAPDH was used as the internal control. The sequences of primers are presented in Table S1.
3
HCC Tissue RNA Extraction and qRT-PCR
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A total of 23 clinical HCC specimens used in this study were histopathologically and clinically diagnosed at the third a liated hospital, Sun Yat-Sen University (Guangzhou, Guangdong, China). All samples were collected from 2017 to 2018 and each HCC tissue has paired non-tumorous tissues. For the use of these tissue in our study, we obtained approval from the Institutional Research Ethics Committee and informed written consent from all participants.
Total RNA extraction and qRT-PCR Total RNA was isolated from tissue specimens using Trizol reagent (Invitrogen, Life Technologies Inc., Germany) according to the manufacturer's instructions. The density and purity of RNA were measured with NanoDrop 2000 (Thermo Fisher Scienti c, USA). cDNA was synthesized by reverse transcription by First Strand cDNA Synthesis Kit for RT-PCR (Roche, Switzerland) according to the manufacturer's instructions. qRT-PCR was performed on LC480 real-time PCR detection system (Bio-Rad, Hercules, CA, USA), and a Roche SYBR FAST Universal qPCR Kit (Roche, Switzerland) was used for gene detection. All primers used for qRT-PCR were designed in primerBank (https://pga.mgh.harvard.edu/primerbank/index.html) and synthesized by BGITech (Beijing, China). GAPDH was used as the internal control. The sequences of primers are presented in Table S1.
Total RNA extraction and qRT-PCR Total RNA was isolated from tissue specimens using Trizol reagent (Invitrogen, Life Technologies Inc., Germany) according to the manufacturer's instructions. The density and purity of RNA were measured with NanoDrop 2000 (Thermo Fisher Scienti c, USA). cDNA was synthesized by reverse transcription by First Strand cDNA Synthesis Kit for RT-PCR (Roche, Switzerland) according to the manufacturer's instructions. qRT-PCR was performed on LC480 real-time PCR detection system (Bio-Rad, Hercules, CA, USA), and a Roche SYBR FAST Universal qPCR Kit (Roche, Switzerland) was used for gene detection. All primers used for qRT-PCR were designed in primerBank (https://pga.mgh.harvard.edu/primerbank/index.html) and synthesized by BGITech (Beijing, China). GAPDH was used as the internal control. The sequences of primers are presented in Table S1.
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