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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.

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2 protocols using rt kits

1

Quantification of Gene Expression in ATDC5 Cells

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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′.
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2

Aortic Tissue RNA Expression Analysis

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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.
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