Ten pairs of CRC and adjacent normal tissues were collected from patients after surgical resection at The First Affiliated Hospital of Anhui Medical University, and the experiments were approved by the Ethics Committee. The samples were stored at -80°C until use. All participants provided signed informed consent. TRIzol reagent (Invitrogen, Carlsbad, CA, USA) was applied to extract total RNA from the collected samples, and the PrimeScript RT kit (Vazyme, Nanjing, China) was used to transcribe extracted RNA into cDNA. The concentrations of all cDNA samples were measured using TB Green Premix Ex Taq II (GenStar, China) with the LightCycler480 System (Applied Biosystems, Waltham, MA, United States). The relative expression levels of signature genes were computed through the 2-ΔΔCt strategy, normalizing to levels of GAPDH. The expression levels between normal and tumor tissues were compared using paired and unpaired t-tests. The primer sequences used for qRT-PCR are presented in Supplementary Table S1
Primescript rt kit
Manufactured by Vazyme
Sourced in China
The PrimeScript RT kit is a reverse transcription kit that enables the conversion of RNA to cDNA, which can then be used for various downstream applications such as PCR and gene expression analysis.
Automatically generated - may contain errors
Lab products found in correlation
Trizol reagent, by Thermo Fisher Scientific (8 mentions)
Iq5 multicolor real time pcr system, by Bio-Rad (3 mentions)
Sybr green, by Vazyme (3 mentions)
Lightcycler480 system, by Thermo Fisher Scientific (2 mentions)
Tb green premix ex taq 2, by GenStar (2 mentions)
Rna easy isolation reagent, by Vazyme (2 mentions)
Gdna eraser, by Vazyme (2 mentions)
Chamq universal sybr qpcr master mix, by Vazyme (2 mentions)
Trizol, by Vazyme (2 mentions)
Sybr qpcr master mix, by Vazyme (2 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
Sybr premix, by Vazyme (1 mentions)
Sybr green master mix, by Vazyme (1 mentions)
Sybr green master mix, by Yeasen (1 mentions)
Cfx96 c1000 thermal cycler, by Bio-Rad (1 mentions)
Aceq universal sybr qpcr master mix, by Vazyme (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Sybr green kit, by Takara Bio (1 mentions)
Fetal bovine serum (fbs), by Solarbio (1 mentions)
Steponeplus real time pcr system, by Thermo Fisher Scientific (1 mentions)
24 protocols using primescript rt kit
1
Comparative Analysis of Colorectal Cancer Transcriptomes
2
Retinal RNA Extraction and qPCR Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
TRIzol (Invitrogen, Carlsbad, CA, United States) was applied to extract total RNA from the retinas of EAU mice on the 14th day after injection. The extracted RNA was reverse transcribed into cDNA using the PrimeScript RT kit (Vazyme, China). SYBR premix (Vazyme, China) was used for final quantitative PCR detection. All the primers are listed in Table 1
3
PVN RNA Extraction and Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total PVN RNA was extracted using the RNA‐easy™ Isolation Reagent (Vazyme Biotech Co.). A PrimeScript™ RT kit (Vazyme Biotech Co.) was used for reverse transcription. Relative mRNA level was detected by in a Bio‐Rad iQ5 Multicolor Real‐Time PCR system (Bio‐Rad Laboratories, CA, USA) with SYBR Green (Vazyme Biotech Co.). The 2−ΔΔCt method was used to calculate the relative targeted gene expression and normalized to GAPDH expression. Primer sequences were listed in Table 1 .
4
Quantitative Gene Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted from tissues or cell lines using TRIzol as directed by the manufacturer (15596018, Thermo). cDNA was then synthesized using the PrimeScript™RT kit (R232-01, Vazyme). The Real-time polymerase chain reaction (RT-PCR) was performed by SYBR Green Master Mix (Q111-02, Vazyme), and the expression levels were counted with the 2−ΔΔCt method. The expression of each mRNA was standardized by the expression level of mRNA GAPDH. All primers were supplied by Tsingke Biotech (Beijing, China), and detailed primer sequences were in Supplementary Table 2
5
RNA Isolation and qRT-PCR Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To isolate RNA, the EZ-press Kit (B0004D, EZB) was employed, followed by reverse transcription utilizing the Prime Script RT kit (R333-01, Vazyme). Amplification was carried out using the SYBR Green Master Mix (11184ES08, YEASEN), using GAPDH as the internal control. To minimize redundancy, the language has been reorganized and revised.
6
RNA Extraction and qPCR Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RNA was prepared via the RNAprep Pure Kit (Magen) with DNase I for reverse transcription using the PrimeScript RT kit (Vazyme). Quantitative PCR was performed with SYBR qPCR Mix (Tsingke) in the CFX 96 C1000 Thermal Cycler (Bio-Rad).
7
Quantitative Gene Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA extraction was executed through the utilization of TRIzol reagent (Life Technologies, Carlsbad, CA, USA), with subsequent complementary DNA (cDNA) synthesis facilitated by a PrimeScript RT kit (Vazyme, Nanjing, China). The concentration of cDNA was quantified using TB Green Premix Ex Taq II (GenStar, Guangdong, China) and a LightCycler480 System (Applied Biosystems, Waltham, MA, United States). Relative expression levels of HOXC6, G0S2, and MX2 were determined employing the 2−ΔΔCt method, with GAPDH serving as the internal control. Differential gene expression across distinct cell lines was assessed utilizing Student’s t−test. Primer sequences are shown in Additional file 1 : Table S2.
8
Quantitative Gene Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA from myocardial tissues or cultured CD4+ T cells was extracted using TRIzol reagent (Invitrogen, Carlsbad, USA) and 1000 ng of RNA was reverse transcribed using the PrimeScript RT kit (Vazyme, Nanjing, China) to generate cDNA. The related primers (Table S1) were purchased from Tsingke (Beijing, China). Data analysis was performed using the 2−ΔΔCT method. The amplified specific cDNA was quantified using AceQ Universal SYBR qPCR Master Mix (Vazyme, Nanjing, China) in a Bio-Rad CFX CONNECT detector system.
9
ATF6 Knockdown Experiments Using siRNA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Two small interfering RNAs (siRNAs) targeting ATF6 genes and their corresponding negative controls (si-NC) were synthesized by Ribobio (Guangzhou, China), The transfection regimen was performed according to the protocol for Lipofectamine 3000(Invitrogen, USA).
Total RNA was extracted from cell lines using TRIzol reagent (15596018, Thermo) and the RNA concentration was standardized. Subsequently, cDNA was synthesized using PrimeScript™RT kit (R232-01, Vazyme). SYBR Green Kit (TaKaRa Biotechnology, Dalian, China) is used for real-time quantitative PCR (qRT-PCR). GAPDH was used as an internal reference.Supplementary Table 1
Total RNA was extracted from cell lines using TRIzol reagent (15596018, Thermo) and the RNA concentration was standardized. Subsequently, cDNA was synthesized using PrimeScript™RT kit (R232-01, Vazyme). SYBR Green Kit (TaKaRa Biotechnology, Dalian, China) is used for real-time quantitative PCR (qRT-PCR). GAPDH was used as an internal reference.
10
Colorectal Cancer Tissue Sampling and qRT-PCR
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In our study, we collected tissue samples from 80 colorectal cancer patients (40 males and 40 females over 65 years old) who came from the Second Affiliated Hospital of Nanchang University (Nanchang, China). Before starting this study, we have informed all patients about the use of human specimens in the study and obtained consent from each patient. All the patients comprehend it and signed paper informed consent.In addition, the ethics committee of the Second Affiliated Hospital of Nanchang University also provided consent for the study. Cell lines were obtained and authenticated from Shanghai Cell Biology (Shanghai, China). All cell lines were tested and were negative for mycoplasma contamination. HCT116, SW480 and LOVO cells were cultured in RPMI 1640 containing 10% fetal bovine serum (FBS) (Solarbio, Beijing, China) and maintained at 37 °C in a humidified atmosphere with 5% CO2. In this experiment, we used TRIzol Reagent (Thermo Fisher, New York, USA) to extract the total RNA from tissue samples and cultured cells and utilized the PrimeScript RT kit (Vazyme, Nanjing, China) and SYBR Premix Ex Taq (China, Chengdu Atomic Energy) to perform quantitative RT-PCR. The following primer sequences were used: KLHL22 Forward) 5ʹ-AAGGGATCTTGTGGCTGT-3ʹand(reverse) 5ʹ-TGTTGTTCACGCAGTGTGG-3ʹ.
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
Revolutionizing how scientists
search and build protocols!