RNA was purified using kit columns (NucleoSpin® RNA, TAKARA, Tokyo, Japan) according to the manufacturer’s instructions. DNA-free RNA was reverse transcripted using GoScript TM reverse transcriptase and random primer (Promega, Madison, USA), according to manufacturer’s instructions. The qRT-PCR was performed by SYBR Premix Ex TaqTM II (Takara, Shiga, Japan) and the specific primers (Table S1 ) on a Thermal Cycler Dice System 2 (Takara). Gene expression of each target gene was calculated using the 2ΔΔCt method (Hamasaki et al., 2019 , Hamasaki et al., 2020 (link)).
Thermal cycler dice system 2
Manufactured by Takara Bio
Sourced in Japan
The Thermal Cycler Dice System 2 is a laboratory instrument designed for DNA amplification. It provides precise temperature control and cycling for PCR (Polymerase Chain Reaction) and other thermal cycling applications.
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Lab products found in correlation
3 protocols using thermal cycler dice system 2
1
RNA Extraction and qRT-PCR Analysis
2
RNA Extraction and qRT-PCR Analysis
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Equal numbers of cells were lysed using TRIzol Reagent (Invitrogen, Waltham, MA), and chloroform (Wako) was added for phase separation and RNA purification. RNA was purified from the aqueous layer using NucleoSpin RNA (Takara, Shiga, Japan) and reverse transcribed using a GoScript Reverse Transcriptase kit (Promega, Madison, WI). The quantitative real-time PCR was performed by SYBR Premix Ex TaqTM II (Takara) on a Thermal Cycler Dice System 2 (Takara) with the specific primers listed in Table 2. Gene expression of each target gene was calculated using the 2 DD threshold cycle (C T ) method. 21
3
Quantitative Real-Time PCR for Gene Expression
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First-strand cDNA was synthesized from purified RNA (0.5 µg) of each sample using GoScript Reverse Transcriptase kit (Promega, Madison, USA) to cDNA. The qRT-PCR was performed using SYBR_ Premix Ex Taq II (Takara, Shiga, Japan) with specific primers (Table S7 ) on a Thermal Cycler Dice System 2 (Takara). Gene expression of target gene was calculated after normalizing to the expression of GAPDH housekeeping gene using the 2ΔΔCT method.
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