Rna pre pure cell bacteria kit

The fluorescence analysis was performed as described previously32 . Briefly, total RNA was extracted from tumours using an RNA pre-pure Cell/Bacteria Kit (Tiangen Biotech, Beijing, China) and reverse-transcribed into cDNA using a FastQuant RT Kit (Tiangen Biotech, China) according to the manufacturer’s instructions. Real-time PCR for the indicated genes was detected on an LC480 (Roche, Basel, Germany) with a SYBR Green Master Mix Kit (Tiangen Biotech, Beijing, China) following the manufacturer’s instructions. The primers used in the experiments are described in Table S1. β-actin mRNA served as an internal reference, and the results were normalized and are shown as the ratio to the control group.

The total RNA from PC12 was extracted using the RNA pre-pure Cell/Bacteria Kit (Tiangen Biotech, China). The concentrations of the total RNA were measured using a Gene Quant 100 Spectrophotometer (General Electric Company, Germany). Five micrograms of total RNA were used to generate complementary DNA using an anchored oligo (dT)₁₈ primer and EasyScript Reverse Transcriptase enzyme (Transgene, China). The cDNA was subsequently used as a template for real-time PCR using SYBR Green Master Mix (Tiangen Biotech, China) according to the manufacturer’s instructions. The primer sequences of RB1 were as follows (NCBI Reference Sequence: NM_009029.2): forward: 5′-TGCTGAAGGCGGCAATCCCC-3′; reverse: 5′- CGAGTCAGGTGTCCCGAGGGT-3′.