Total RNA from the HUVECs was isolated using the RNeasy mini kit (Qiagen, Hilden, Germany), and first strand of the cDNA was synthesized using a Transcriptor First Strand cDNA synthesis kit (Bio-Rad). Quantitative reverse transcription-PCR was performed using iTaq universal SYBR Green supermix (Bio-Rad) according to the manufacturer’s protocol. Primers used were as follows. ICAM-1 sense: 5′-AGG GTA AGG TTC TTG CCC AC -3′; ICAM-1 antisense: 5′-TGA TGG GCA GTC AAC AGC TA -3′, VCAM-1 sense: 5′-GTC TCC AAT CTG AGC AA -3′; VCAM-1 antisense: 5′-TGG GAA AAA CAG AAA AGA GGT G -3′, GAPDH sense: 5′-TTG ATG GCA ACA ATC TCC AC -3′; GAPDH antisense: 5′-CGT CCC GTA GAC AAA ATG GT -3′.
Transcriptor first strand cdna synthesis kit
Manufactured by Bio-Rad
Sourced in Germany, United States
The Transcriptor First Strand cDNA Synthesis Kit is a laboratory product designed for the reverse transcription of RNA into complementary DNA (cDNA). The kit contains the necessary reagents and enzymes to perform this process, which is a fundamental step in various molecular biology and gene expression analysis techniques.
Automatically generated - may contain errors
Lab products found in correlation
Rneasy mini kit, by Qiagen (1 mentions)
Itaq universal sybr green supermix, by Bio-Rad (1 mentions)
Real time pcr primers, by Thermo Fisher Scientific (1 mentions)
Thermo scientific nanodrop 2000 spectrophotometer, by Thermo Fisher Scientific (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Sybr green master mix, by Bio-Rad (1 mentions)
2 protocols using transcriptor first strand cdna synthesis kit
1
Quantitative RT-PCR Analysis of ICAM-1 and VCAM-1 Expression
2
Quantifying Liver Fibrosis Gene Expression
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RNA was isolated from liver fibrosis tissues or the HSCs by using Trizol Reagent (Invitrogen) according to the manufacturer’s instructions. Total RNA quality and density were assessed by using the Thermo Scientific NanoDrop 2000 Spectrophotometer (Thermo Fisher Scientific, United States). To qualify miRNA or mRNA, RNA was reverse transcribed to cDNA by using transcriptor first-strand cDNA synthesis kit (Bio-Rad, United States) following the manufacturer’s protocols. After that, real-time PCR was carried out in a detection system with SYBR-Green Master Mix (Bio-Rad). Real-time PCR primers were acquired from Invitrogen. For the miR-145 expression analysis, the one-step miRNA real-time PCR Detection Kit (biomics, Nantong, Jiangsu, China) was used according to the manufacturer’s protocols. U6 small nuclear was used as endogenous control. All of measurements were performed three times and GAPDH was used as an endogenous control for mRNA expression. The following primers of genes were used:
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