First strand cdna synthesis supermix

Reverse transcription of total RNA extracted from 10 lesions and adjacent healthy lung tissues into cDNA was performed using First-Strand cDNA Synthesis SuperMix (Takara, Dalian, China). Quantitative real-time PCR (qRT-PCR) then was performed with specific primers using SYBR Green PCR kit (Invitrogen), according to the manufacturer’s protocol. Expression of GAPDH was used as internal standard for lncRNAs and mRNAs, U6 was used as internal standard for miRNAs. Results were analyzed using 2^−ΔΔCt method. Primer sequences which designed by sagan corporation are shown in Supplemental Table 1. A P < 0.05 was considered significant difference.

Total RNA was extracted from the myocardial tissues of the HF and control groups using a TRIzol extraction kit (Invitrogen, Carlsbad, CA, USA). Reverse transcription of RNA into cDNA was then conducted using the First-Strand cDNA Synthesis SuperMix (Takara, Dalian, China). Quantitative real-time PCR (qRT-PCR) was performed using specific primers (Table 2) according to the SYBR Green PCR Kit (Invitrogen, Carlsbad, CA, USA) according to the protocol of the manufacturer. About 35 PCR cycles were used for the amplification. GAPDH mRNA expression level was used as an internal standard, and the results were analyzed using the 2^−ΔΔCt method.