Total RNA was extracted using the RNAeasy™ Animal RNA Isolation Kit with Spin Column (Beyotime, China) according to manufacturer’s instructions. Total RNA was reversely transcribed amplified using the BeyoFast™ SYBR Green One-Step qRT-PCR Kit (Beyotime, China). All real-time PCR primers were listed as following: AURKA, 5′-CTAACGGCTGAGCTCTTGGA-3′ and 5′-GAACCGACAGGGGACTTGAC-3′. CCNB1, 5′-ACCTTTGCACTTCCTTCGGA-3′ and 5′-TGTTCTTGACAGTCCATTCACCA-3′. CDK1, 5′-GCCCTTTAGCGCGGATCTAC-3′ and 5′-AGGAACCCCTTCCTCTTCACT-3′. TOP2A, 5′-CCGTCACCATGGAAGTGTCA-3′ and 5′-TGTCTGGGCGGAGCAAAATA-3′. CYP2B6, 5′-CCTCAACCTCAACACGCTCT-3′ and 5′-TTTGGCTCGGTCATGAAGCT-3′. CYP2C9, 5′-ACCAGCTGTGCTTCATTCCT-3′ and 5′-GCACAGTGAAACATAGGAAACTCTC-3′. CYP3A4, 5′-GCTTTCCTGCACATTAAGGAGAA AT-3′ and 5′-ATGGGCAAAGTCACAGTGGAT-3′. GAPDH, 5′-AGCCTCAAGATCATCAGC-3′ and 5′-GAGTCCTTCCACGATACC-3′. Relative mRNA expression levels of target genes were normalized by comparing to GAPDH, and calculated using the 2 − ΔΔCt method [41 (link)]. Expression data of selected genes were processed by GraphPad Prism 9, and analyzed by Student’s t-test. P-value less than 0.05 was regarded as the cutoff value of statistical significance.
Beyofast sybr green one step qrt pcr kit
Manufactured by Beyotime
Sourced in China
BeyoFast™ SYBR Green One-Step qRT-PCR Kit is a real-time reverse transcription PCR (qRT-PCR) kit that uses SYBR Green I as the fluorescent dye. It is designed for the quantitative detection of RNA targets.
Automatically generated - may contain errors
Lab products found in correlation
Rnaeasy animal rna isolation kit with spin column, by Beyotime (2 mentions)
Trizol reagent, by Thermo Fisher Scientific (2 mentions)
Trizol, by Thermo Fisher Scientific (2 mentions)
Prism 9, by GraphPad (1 mentions)
Beyozol, by Beyotime (1 mentions)
Tanon 5200 imaging system, by Shanghai Tanon Science & Technology (1 mentions)
Cfx96 real time pcr detection system, by Bio-Rad (1 mentions)
Abi quantstudio 6 flex, by Thermo Fisher Scientific (1 mentions)
Rnaeasy animal rna isolation kit, by Beyotime (1 mentions)
Microrna reverse transcription kit, by Takara Bio (1 mentions)
Abi 7500 fast pcr system, by Toyobo (1 mentions)
Trizol reagent, by Beyotime (1 mentions)
Universal genomic dna purification mini spin kit, by Beyotime (1 mentions)
Sybr green pcr master mix, by Toyobo (1 mentions)
Beyort 3 first strand cdna synthesis kit, by Beyotime (1 mentions)
Stratagene mx3000p instrument, by Agilent Technologies (1 mentions)
Trizol reagent, by Takara Bio (1 mentions)
Western blocking buffer, by Beyotime (1 mentions)
C11 bodipy581 591, by Thermo Fisher Scientific (1 mentions)
Pvdf membrane, by Beyotime (1 mentions)
16 protocols using beyofast sybr green one step qrt pcr kit
1
Quantitative Real-Time PCR Analysis
2
RNA Extraction and qPCR Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Extraction of total RNA was performed with TRIzol reagent (Life Technologies) following the manufacturer’s instructions, and the extracted RNA was then quantified. Next, qPCR was performed using a BeyoFast™ SYBR Green One-Step qRT-PCR Kit (Beyotime, D7268S). The expression of MAP3K15 was normalized to that of GAPDH. Relative expression levels were calculated with the 2−∆∆Ct method.
3
Profiling HDAC6 Splice Variants
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA from HCT116 cells treated with or without compound 6d for 4 h and 8 h was extracted using Beyozol (R0011, Beyotime) according to the manufacturer’s instructions. The genomic sequence of HDAC6 spanning exons 26 to 29 was amplified using the forward primer 5′-GGGGGATCCGGGGCCTCAGAATCTCAG-3′ and the reverse primer 5′-GGGGCTCGAGAACAGCTTGTACTTTATT-3′. RT-PCR was performed using BeyoFast™ SYBR Green One-Step qRT-PCR Kit (D7268S, Beyotime) according to manufacturer’s instructions, and then the PCR products were assessed to detect the relative abundance of different splice forms by applying to 2% agarose gels. The results were analyzed using the Tanon 5200 Imaging System (Tanon Science & Technology Co., Ltd., Shanghai, China).
4
Quantifying mRNA Expressions of Key Genes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To test the mRNA expressions of DNMT1, TGFβ1, PTEN, and TSC2, quantitative real-time PCR was conducted. After relevant treatment, cells were harvested in 1.5 mL EP tubes and 1.0 mL of Trizol solution was added for 5 min at room temperature. Then, 0.2 mL chloroform was supplemented into tube and tube was centrifuged at 4°C at 12000rmp for 15 min. Supernatant was discarded and 1 mL of 75% ethanol was added to wash RNA precipitate one time (centrifuged at 7500rmp for 5 min). RNA concentration was determined using a microamount RNA/DNA quantimeter. The relative expressions of DNMT1, TGFβ1, PTEN, and TSC2 were detected using BeyoFast™ SYBR Green One-Step qRT-PCR Kit (Beyotime Biotechnology, Shanghai, China). The reaction conditions were set as: (1) 95°C for 10 min, (2) 40 cycles of 95°C for 5 s and 60°C for 30 s, (3) 94°C for 30 s, 60°C for 90 s, and 94°C for 10 s. Primer's information was displayed in Table 2 . Data were analyzed using 2-△△CT method and β-actin was served as internal control.
5
Quantification of Apelin mRNA Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted using RNAeasy™ Animal RNA Isolation Kit with Spin Column (Beyotime Biotechnology, Shanghai, China). The following primers were used to study the expressions of mouse GAPDH and apelin: mouse GAPDH forward: 5′-CCACTGTGGGCCACTTATACC-3′; mouse GAPDH reverse: 5′-CAGCCTTAGCCGAGCATTG-3′; mouse apelin forward: 5′-GGAATTCGGGACCATGAATCTGAGGCTCTG-3′; and mouse apelin reverse: 5′-ACTTGGCGAGCCCTTCAATC-3′. The qRT-PCR analysis was conducted using BeyoFast™ SYBR Green One-Step qRT-PCR Kit (Beyotime Biotechnology, Shanghai, China) on the platform of a CFX96 Real-Time PCR Detection System (Bio-Rad, Hercules, CA, USA). The relative abundance of apelin was normalized by the level of GAPDH mRNA expression.
6
Quantitative RT-PCR Analysis of Tight Junction Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RNA was extracted from frozen colonic tissue using the RNAeasy™ Animal RNA Isolation Kit and a Spin Column (R0027, Beyotime) according to the manufacturer’s instructions. The contents of occludin, claudin-4, and ZO-1 mRNA were measured by quantitative reverse transcription PCR. Quantitative reverse transcription PCR was performed using the BeyoFast™ SYBR Green One-Step qRT-PCR Kit (D7268S, Beyotime), and relative RNA expression was measured and analyzed using the ABI QuantStudio 6 Flex (Thermo Fisher). Each reaction was performed in triplicate. The relative amount of RNA compared to the internal control was calculated by the 2−ΔΔCt method. GAPDH was used as the internal reference gene. The relative primer sequences are available in Supplementary Table 1 (n = 4 for the AMVN group and n = 5 for the AKK group).
7
Gene Expression Analysis via qRT-PCR
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
TRIzol reagent (Beyotime, Shanghai, China) was used to extract total RNA. BeyoFast™ SYBR Green One-Step qRT‒PCR Kit (Beyotime) was used to detect targeting gene according to the manual. In addition, for miRNAs, MicroRNA Reverse Transcription Kit (Takara Biotechnology, Japan) were used to perform reverse transcription. qRT-PCR was carried out on ABI 7500 fast PCR System (Carlsbad, CA, USA) with a SYBR green PCR Master Mix (TOYOBO, Japan). β-actin and U6 applied as internal references for mRNAs and miRNAs. The relative expressions were calculated with 2–ΔΔCT method. Moreover, genomic DNA (gDNA) was extracted by a Universal Genomic DNA Purification Mini Spin Kit (Beyotime) based on the manufacturer’s protocol. All primers were synthesized by GenePharma (Shanghai, China) and are listed in Table S2 .
8
Quantifying Osteogenic Marker Genes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Expression levels of osteogenic marker genes and cytokine receptor genes in LF cells were assessed by quantitative real‐time polymerase chain reaction (qRT‐PCR). Total RNA was isolated using Trizol (Invitrogen, Carlsbad, CA). Reverse transcription and qPCR were performed using a BeyoFast™ SYBR Green One‐Step qRT‐PCR Kit (D7268; Beyotime, Shanghai, China) according to the manufacturer's instructions on a BioRad IQ5 system (BioRad, Hercules, CA). Values were normalized to glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) using the 2−ΔΔCt method. Primer sequences are provided in Table 2 .
9
Quantification of Lung iNOS mRNA by qRT-PCR
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The mRNA level of iNOS in lung tissue was determined by quantitative real-time polymerase chain reaction (qRT-PCR) as described previously (Rungsung et al. 2018 (link)). In brief, total RNAs were isolated from the collected lung tissues with the help of Trizol reagent (TAKARA, Beijing, China) and were finally dissolved in diethylpyrocarbonate (DEPC; MACKLIN, Shanghai, China)-treated ddH2O. Afterwards, the first strand of cDNA synthesis was fulfilled using a BeyoRT™ III First Strand cDNA Synthesis Kit (Beyotime Biotechnology, Shanghai, China). Then, the iNOS expression was determined using BeyoFast™ SYBR Green One-Step qRT-PCR Kit (Beyotime Biotechnology) with a Stratagene Mx3000P instrument (Agilent Technologies, California, USA). GAPDH was used as the internal control, and the 2-ΔΔCt method was used to calculate iNOS expression. The sequences were as follows: inducible nitric oxide synthase (iNOS) (Forward) 5′-GGTGCTATTCCCAGCCCAA-3′, (Reverse) 5′-AGTCACATGCAGCTTGTCCA-3′; GAPDH (Forward) 5′-AGACAGCCGCATCTTCTTGT-3′, (Reverse) 5′-CCGATACGGCCAAATCCGTT-3′.
10
Ferroptosis Regulation in Cancer Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DuIbecco’s modified eagIe’s medium (DMEM) and fetal bovine serum (FBS) were obtained from Gibco (Grand Island, NY, USA). D-glucose, PD (purity ≥ 98% HPLC; Figure 2(a) ), Thiazolyl Blue Tetrazolium Bromide (MTT), dimethyl sulfoxide (DMSO), and PBS were purchased from Sigma-Aldrich (St. Louis, MO, USA). Erastin and deferasirox (DFX) were purchased from MedChemExpress (Monmouth Junction, NJ, USA). Small interfering RNA (si)-GPX4 1#, si-GPX4 2#, and its negative control (si-nc) were acquired from GenePharma (Shanghai, China). Lipofectamine 3000, TRIzol, and C11-BODIPY (581/591) were acquired from Invitrogen (Carlsbad, CA, USA). The lactate dehydrogenase (LDH) assay kit was purchased from Jiancheng (Nanjing, China). Reduced and oxidized glutathione (GSH and GSSG, respectively) assay kit, lipid peroxidation malonaldehyde (MDA) assay kit, BCA protein assay kit, PVDF membranes, western blocking buffer, BeyoECL Plus, terminal deoxynucleotidyl transferase biotin-dUTP nick-end labeling (TUNEL) apoptosis assay kit, and BeyoFast SYBR Green one-step qRT-PCR kit were purchased from Beyotime (Shanghai, China). The iron assay kit, primary antibodies (anti-ACSL4, anti-TFR1, anti-FTH-1, anti-SLC7A11, anti-GPX4, and anti-GAPDH), and HRP-conjugated secondary antibody were purchased from Abcam (Cambridge, MA, USA).
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!