Total RNA was extracted from OS tissues, corresponding para-tumour tissues, and OS cells using the Trizol reagent (Invitrogen, USA) according to the manufacturer’s protocol. RNA was reverse-transcribed into cDNAs using the Hifair™ III 1st Strand cDNA Synthesis Kit (Yeasen, China) or miRNA First Strand cDNA Synthesis Kit (Sangon Biotech, China). The mRNA or lncRNA level was assessed the using Hieff™ qPCR SYBR® Green Master Mix Kit (Yeasen, China), and the miRNA level was assessed using the miRNAs qPCR Kit (Sangon Biotech, China) and ABI7500 system (Applied Biosystems, USA). GAPDH or small nuclear RNA U6 served as the internal control. The relative expression levels were calculated using the 2-ΔΔCt method. The primers sequences are listed in Supplementary Table 1 .
Mirnas qpcr kit
Manufactured by Sangon
Sourced in China
The MiRNAs qPCR Kit is a laboratory equipment designed for the quantitative detection and analysis of microRNAs (miRNAs) using real-time PCR technology. The kit provides a standardized and reliable solution for the accurate quantification of miRNA expression levels in various biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Mirna first strand cdna synthesis kit, by Sangon (3 mentions)
Trizol reagent, by Thermo Fisher Scientific (3 mentions)
Abi 7500 system, by Thermo Fisher Scientific (2 mentions)
Hifair 3 1st strand cdna synthesis kit, by Yeasen (1 mentions)
Hieff qpcr sybr green master mix kit, by Yeasen (1 mentions)
Trizol reagent, by Merck Group (1 mentions)
Sybr premix ex taq, by Takara Bio (1 mentions)
Abi 7300 real time pcr detection system, by Thermo Fisher Scientific (1 mentions)
Cdna synthesis kit, by Takara Bio (1 mentions)
Bulge loop mirna primer set, by RiboBio (1 mentions)
4 protocols using mirnas qpcr kit
1
Quantitative Analysis of RNA Expression
2
Quantifying miR-326 and TNFSF14 mRNA Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To detect the expression of miR-326 and TNFSF14 mRNA, qRT-PCR assay was performed. Total RNA was collected from ASMCs using Trizol reagent (Sigma-Aldrich, St Louis, Missouri). The extracted RNA was used to perform reverse transcription to generate cDNA by PrimeScript RT Master Mix Kit (Sigma-Aldrich) or miRNA First Strand cDNA Synthesis Kit (Sigma-Aldrich). qPCR was then carried out using SYBR Premix Ex Taq (Takara, Dalian, China) and miRNAs qPCR Kit (Sangon Biotech, China). GAPDH and U6 were used as controls. The primers sequences were listed as follows: miR-326: forward: 5′-AACAATCCTCTGGGCCCTTC-This website utilizes technologies such as cookies to enable essential site functionality, as well as for analytics, personalization, and targeted advertising. To learn more, view the following link: Privacy Policy 3′, reverse: 5′-GTCGTATCCAGTGCAGGGT-3′; TNFSF14: forward: 5′-GGTCTCACGAGGTCAACCCA-3′, reverse: 5′-CCAACAGCTCCAGCTCCTC-3′. The relative miR-326 and TNFSF14 mRNA expression were analyzed by the 2 -ΔΔCt method.
3
Quantitative Analysis of mRNA and miRNA Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted from cultured CFs and left atriums using Trizol reagent (Invitrogen, CA), according to the manufacturer's instructions. Total RNA (500 ng for mRNA or 1000 ng for miRNA) from each sample was subjected to reverse transcription according to the instructions of the cDNA Synthesis Kit (TaKaRa, China) and miRNA First Strand cDNA Synthesis Kit (Sangon, China). The expression levels of mRNA and miR‐30c were analysed according to the instructions of the mRNA SYBR qPCR kit (TaKaRa, China) and miRNAs qPCR Kit (Sangon, China) using the ABI‐7300 Real‐Time PCR Detection System (Applied Biosystems, USA). The mRNA primer sequences used in the study are listed in Table 1 . The bulge‐loop™ miRNA Primer Sets (one RT primer and a pair of qPCR primers) specific for miR‐30c and U6 were purchased from RiboBio (Guangzhou, China). The levels of mRNA and miR‐30c were normalized to β‐Actin and U6, respectively. Briefly, the cycle threshold (CT) values of each targeting gene were subtracted from the CT values of the housekeeping gene, referring as ▵ct. Target gene ▵▵ct was determined as ▵ct of target gene minus ▵ct of control. The relative expression levels of all genes were calculated basing on the 2−ΔΔct. All samples were assayed in triplicate.
4
Quantification of miRNA Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total cellular RNA was extracted using the Trizol reagent (Invitrogen, USA) and RNAs were reverse-transcribed into cDNAs using the miRNA First Strand cDNA Synthesis Kit (Sangon Biotech, China). All miRNA levels were assessed using the miRNAs qPCR Kit (Sangon Biotech, China) and the ABI7500 system (Applied Biosystems, USA). Small nuclear RNA U6 was used as an internal reference. There were three replicates for each sample, and the cycle threshold (Ct) values were averaged. The relative expression of each miRNA was calculated using the 2−△△Ct method. Supplementary Table 1
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!