Total RNA was isolated from Arabidopsis seedlings using Takara RNAiso Plus reagent (TAKARA, http://www.takara-bio.com/ ). Quantitative real-time PCR was performed using the TB Green® Premix Ex Taq™ Tli RNase H Plus (TAKARA, http://www.takara-bio.com/ ), following the manufacturer’s instructions. The relative abundance of each cDNA was normalized against TUB8 (AT5G23860) or ACT1 (AT2G37620). The sequences of gene-specific primers are provided in Supplemental Table 1 . Primers used to detect HY5 (AT5G11260) transcripts were the same as reported previously (Favory et al. 2009 (link)).
Takara rnaiso plus reagent
Manufactured by Takara Bio
Sourced in United States, Japan
Takara RNAiso Plus reagent is a complete solution for the isolation of total RNA from various biological samples. It is designed to effectively extract and purify high-quality RNA for downstream applications.
Automatically generated - may contain errors
Lab products found in correlation
Tb green premix ex taq tli rnaseh plus, by Takara Bio (1 mentions)
Pikoreal 96 real time pcr system, by Thermo Fisher Scientific (1 mentions)
Primescript rt reagent kit, by Takara Bio (1 mentions)
Iscript cdna synthesis kit, by Bio-Rad (1 mentions)
Itaq universal sybr green supermix, by Bio-Rad (1 mentions)
3 protocols using takara rnaiso plus reagent
1
Quantitative RT-PCR for Arabidopsis Transcripts
2
Quantitative Real-Time PCR Analysis
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Inguinal white adipose tissue and 3T3‐L1 cells were homogenized in TAKARA RNAisoPlus reagent (Takara), and single standard complimentary deoxyribonucleic acid (cDNA) was synthesized by using Prim eScript RT Reagent Kit (Takara, USA). Quantitative real‐time PCR was performed with Thermo Scientific PikoReal 96 Real‐Time PCR System (Waltham, USA). Each reaction was performed in duplicate and the value of the gene of interest was normalized to mouse GAPDH or RPS18 expression. The comparative threshold cycle (CT) method was used to calculate the relative expression. The specific primers are shown in Table S1 .
3
RNA Isolation and qPCR Protocol
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Isolation of total RNA pool from cells was performed using TAKARA RNA-iso Plus reagent (Cat No.9109; Takarabio, Japan). First strand cDNA synthesis was performed using iscript™ cDNA Synthesis kit (Cat No. 1708891, Bio-Rad) and subsequent quantitative real-time PCR (q-PCR) using iTaq Universal SYBR Green Supermix (Cat No. 1725120, Bio-Rad), as per earlier discussed protocol.51 (link) The primer sequences are given in Table S1 .
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