Sham or I/R hearts (n = 6 per group) were harvested. Total RNA was isolated from the border zone of the heart with TRIzol reagent (Takara). Equal amounts (1–2 μg) of total RNA and RT Enzyme mix (Takara) were used to synthesize cDNA. The mRNA expression of the target genes was analysed with a PCR thermocycler (Applied Biosystems) as previously described [21 (link)]. The data were normalized to GAPDH expression levels. All PCR primer sequences for each gene used are provided in Table 1 .
Rt enzyme mix
Manufactured by Takara Bio
The RT Enzyme mix is a blend of reverse transcriptase and RNase inhibitor enzymes designed for the efficient conversion of RNA into cDNA. The core function of this product is to facilitate the reverse transcription of RNA samples into complementary DNA (cDNA) for downstream applications such as PCR and other molecular biology techniques.
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Lab products found in correlation
3 protocols using rt enzyme mix
1
Assessing Gene Expression in Myocardial Infarction
2
Gene Expression Analysis by RT-qPCR
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We extracted total RNA utilizing a TRIzol RNA extraction kit (Invitrogen, Carlsbad, CA, United States). The first-strand cDNA was synthesized using an PrimeScript RT Enzyme Mix (Takara RR036A). RT-qPCR was performed using a LightCycler LC480 instrument (Roche) according to the procedures of the manufacturer. 18S was used as internal control to normalize tests. Primer sequences are listed in Supplementary Table 1 .
3
Quantification of Cardiac Gene Expression
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Total RNA was purified from heart border zone tissues obtained from sham and I/R mice (n = 6) with TRIzol (#9109, TaKara). The cDNA was synthesized with total RNA (1 μg) using reverse transcription (RT) enzyme mix (#RR047A, TAKARA). The mRNA levels of Bax, Bcl-2, interleukin (IL)-1β, IL-6, NOX2, NOX4, TNF-α, NQO1, Gclc, SOD1, and Prdx1 were measured by PCR, which was performed with a thermocycler (Bio-Rad), and were normalized to the level of GAPDH. The primers are provided in Table S2 .
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