Rna amplification kit

Then, 400 μl lysis buffer, three grinding beads, and 50 mg hepatic tissue were mixed in a grinding tube and subsequently broken with a homogenizer (5000 rpm, 20 seconds). The tube was then centrifuged (12000 rpm, 5 min, 4°C), and 300 μl of supernatant was placed in a new 1.5 ml RNase-free tube. Next, RNA was extracted using a Universal RNA Extraction kit. The reverse transcriptase reaction was performed for 15 min at 37°C and 5 seconds at 85°C. Finally, a 10 μl PCR mix was applied, including 2 μl cDNA, 5 μl 2× TB green, 0.4 μl forward primer (10 μm), 0.4 μl reverse primer (10 μm), and 2.2 μl ddH₂O.
The Universal RNA Extraction kit (cat: 9767; lot: AJ82351A), reverse transcription (RT) kit (cat: RRO36A; lot: A140832A), and RNA amplification kit (cat: RR820A; lot: A140778A) were purchased from Takara.
We designed primer sequences according to the mRNA sequences obtained from the NCBI database (https://www.ncbi.nlm.nih.gov) and PrimerBank (https://pga.mgh.harvard.edu/primerbank/), synthesized by Sangon Biotech, as shown in Table 1.

The alphalipoic acid injection (Yabao Pharmaceutical Group Co. Ltd.), nab-paclitaxel injection (Fresenius Kabi USA, LLC), and normal saline injection (Tianrui Pharmaceutical, Zhejiang, China) were obtained from Zhongshan Hospital of Fudan University (Shanghai, China). The mRNA extraction kit, cDNA extraction kit, RNA amplification kit, primer design, and synthesis were provided by Takara (Takara Bio Inc., Shiga, Japan). Protein antibodies were purchased from Abcam Inc. (Cambridge, MA, USA).

Erlotinib Hydrochloride Tablets (150 mg erlotinib in each tablet) were provided by Roche Ltd. Due to their insolubility in water, they were made into suspension with 0.5% sodium carboxymethyl cellulose. FBS, Trypsin enzyme and high glucose DMEM medium were purchased from HyClone. mRNA extraction kit, cDNA extraction kit, RNA amplification kit, primer design and synthesis were provided by Takara. Protein antibody was purchased from Cell Signaling.