Total RNA was extracted from H9C2 cell with the use of TRIzol reagent (Invitrogen Life Technologies, Carlsbad, CA, USA). Isolated RNA (10 µg) was reverse-transcribed using the RevertAid First Strand cDNA Synthesis kit (therm k1622; Thermo Scientific, Waltham, MA, USA). PCR amplification was conducted in a total volume of 25 µl (QPS-201; Toyobo, Osaka, Japan): cDNA 2.5 µl, F (5 pmol/ml) 2 µl, R (5 pmol/ml) 2 µl, THUNDERBIRD™ SYBR Green pPCR Mix 12.5 µl and H2O 6 µl, using an Applied Biosystems 7300 Real-Time PCR System (Applied Biosystems Life Technologies, Foster City, CA, USA). The primer sequence for rat GADPH was as follows: F, 5′-CGCTAACATCAAATGGGGTG-3′; and R, 5′-TTGCTGACAATCTTGAGGGAG-3′. GAPDH was used as an internal control. Cycling conditions were: 95°C for 1 min (95°C for 15 sec, 58°C for 20 sec, 72°C for 20 sec) for 40 cycles, and 82°C for 10 min. Data were calculated using the 2−ΔΔCt method.
Qps 201
Manufactured by Toyobo
Sourced in Japan
The QPS-201 is a laboratory equipment product manufactured by Toyobo. It is designed to perform quantitative protein assays. The core function of the QPS-201 is to accurately measure the concentration of proteins in a sample.
Automatically generated - may contain errors
Lab products found in correlation
Revertaid first strand cdna synthesis kit, by Thermo Fisher Scientific (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
7300 real time pcr system, by Thermo Fisher Scientific (1 mentions)
Dp431, by Tiangen Biotech (1 mentions)
Fsq 201, by Toyobo (1 mentions)
Revertra ace qpcr rt master mix, by Toyobo (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
Gdna remover, by Toyobo (1 mentions)
Fsq 301, by Toyobo (1 mentions)
4 protocols using qps 201
1
Quantitative RT-PCR Analysis of GAPDH in H9C2 Cells
2
Quantitative RT-PCR Analysis of ICH
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After obtaining brain tissue from mice at different ICH time points (n=6), total RNA was extracted using a total RNA extraction kit (DP431, TIANGEN), and the concentration and purity were determined using a NanoVue Plus Micro-Volume UV-Vis spectrophotometer. The total RNA was then reverse transcribed into cDNA using a reverse transcription kit (FSQ-201, TOYOBO), and the concentration and purity of the cDNA were determined. Finally, 20μL of amplification reaction mix (QPS-201, TOYOBO) was prepared and the samples were introduced into an ABI StepOne Real-Time PCR system. The primer sequences are listed in Table 2 . The ΔΔCT method was used to calculate the results. The data obtained were analyzed by one-way ANOVA. Tukey multiple comparison was used to test for statistical significance between groups.
3
Quantification of miRNA-124a Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The operation was performed as previous described [22 (link)]. In brief, total RNA was extracted from the MH7A cells and exosomes by Trizol (Thermo Scientific). 1 μg of RNA was reverse transcribed into cDNA by a ReverTra Ace qPCR RT Master Mix with gDNA Remover (TOYOBO) following the manufacturer’s protocol. Then, we performed amplification for 40 cycles (TOYOBO, QPS-201). The level of miRNA-124a was normalized to RNU6, and 2−ΔΔCt method was used to evaluated the relative expression of gene. The primers sequence was listed as follow: U6: (5′-GCTTCGGCAGCACATATACTAAAAT-3′), miRNA-124a:(5′-TAAGGCACGCGGTGAATGCC-3′).
4
Quantification of Multidrug Resistance Genes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted from Huh7 P and Huh7 LR cells using TRIzol reagent, and cDNA synthesis was conducted using a reverse transcription kit (Toyobo FSQ 301, Japan) following the manufacturer’s protocol. Subsequently, qRT-PCR was performed in a total volume of 20 μL, containing Milli-Q water (2 μL), c-DNA (6 μL), forward and reverse primers (2 μL), and q-PCR mix (10 μL; Toyobo QPS-201, Japan). The primers used in this study were manufactured by Ruibiotech (Beijing, China) with the following sequences: β-actin forward: 5′-ATCGTCCACCGCAAATGCTTCTA-3′ and reverse: 5′-AGCCATGCCAATCTCATCTTGTT-3′, MDR1 forward: 5′-GGGAGCTTAACACCCGACTTA-3′ and reverse: 5′-GCCAAAATCACAAGGGTTAGCTT-3′, and BCRP forward: 5′-GCCACAGAGATCATAGAGCCT-3′ and reverse: 5′-TCACCCCCGGAAAGTTGATG-3′. The results were normalised to β-actin expression and are presented as relative mRNA expression levels.
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!