Total RNA from ATDC5 cells was extracted with TRIzol® (Thermo Fisher Scientific, Inc.) according to the supplier's protocol. Next, RT kits (Takara Bio, Inc.) were used to reverse transcribe RNA into cDNA according to the manufacturer's protocol. Subsequently, 50 ng cDNA was used for qPCR using TB Green® Fast qPCR Mix (Takara Bio, Inc.) and an Applied Biosystems 7500 Real-Time PCR System (Thermo Fisher Scientific, Inc.). The following thermocycling conditions were used for qPCR: 95°C for 2 min; followed by 40 cycles of 95°C for 20 sec and 65°C for 40 sec. The results were analyzed using the 2−ΔΔCq method (18 (link)) with Gapdh as the internal reference gene. The primers were as follows: FABP4 forward, 5′-TTCCTTCAAACTGGGCGTGG-3′ and reverse, 5′-GCCTTTCATAACACATTCCACC-3′; matrix metalloproteinase (Mmp)3 forward, 5′-TCCCACATCACCTACAGGATTG-3′ and reverse, 5′-CAGGCCCATCAAAAGGGACA-3′; Mmp9 forward, 5′-CAGCCGACTTTTGTGGTCTTC-3′ and reverse, 5′-CGGTACAAGTATGCCTCTGCCA-3′; Mmp13 forward, 5′-GGAGCCCTGATGTTTCCCAT-3′ and reverse, 5′-GTCTTCATCGCCTGGACCATA-3′; ADAM metalloproteinase with thrombospondin type 1 motif 4 (Adamts-4) forward, 5′-CAAGCATCCGAAACCCTGTC-3′ and reverse, 5′-ACACAGGTCCTGCCGGG-3′; and Gapdh forward, 5′-GGGTCCCAGCTTAGGTTCATC-3′ and reverse, 5′-CCAATACGGCCAAATCCGTTC-3′.
Rt kits
Manufactured by Takara Bio
Sourced in Japan
RT kits are products designed for reverse transcription, a process that converts RNA into complementary DNA (cDNA). These kits provide the necessary reagents and protocols to perform this fundamental step in various molecular biology applications, such as gene expression analysis, RNA sequencing, and cDNA synthesis.
Automatically generated - may contain errors
Lab products found in correlation
Applied biosystems 7500 real time pcr system, by Thermo Fisher Scientific (2 mentions)
Trizol, by Thermo Fisher Scientific (2 mentions)
Tb green fast qpcr mix, by Takara Bio (1 mentions)
Trizol reagent, by Takara Bio (1 mentions)
Tb green premix ex taq 2 kit, by Takara Bio (1 mentions)
Lightcycler 480 system, by Roche (1 mentions)
2 protocols using rt kits
1
Quantification of Gene Expression in ATDC5 Cells
2
Aortic Tissue RNA Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The total RNA was extracted from the aortic tissues using Trizol reagent (Takara, Japan) in accordance with the manufacturer's protocols. Total RNA (1 μg) was then converted to cDNA by the RT kits (Takara, Japan). qRT-PCR was performed on a Roche LightCycler480 System (Roche, USA) using a TB Green™ Premix Ex Taq™ II kit (Takara, Japan) to measure and analyze the expression of specific genes including IL-6, IL-2, VCAM-1, ICAM-1, LXRα, ABCA1, and ABCG1. The primer sequences (Sangon Biotech, China) used in this study were listed in Table 1 . The amplification steps were as follows: denaturing at 95°C for 30 s, followed by 40 cycles of PCR reaction at 95°C for 5 s and 60°C for 20 s, then with a final dissociation stage at 95°C for 5 s, 60°C for 1 min, and 95°C for 0 s. The level of GAPDH was used as an internal control.
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!