Dl2000 dna marker

Rutin, hesperidin, myricetin and quercetin (98% HPLC grade purity) were purchased from Chengdu Alfa Biotechnology Co., Ltd. (Chengdu, China). HPLC grade methanol and glacial acetic acid were obtained from Fuchen (Tianjin) Chemical Reagent Co. Ltd. (Tianjin, China). Anhydrous ethanol (analytical pure AR) were obtained from Tianjin Hengxing Chemical Reagent Manufacturing Co., Ltd. (Tianjin, China).
The assay kits for alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and triglyceride (TG) were procured from Shanghai Fuxing Long March Medical Science Co., Ltd. (Shanghai, China). Superoxide dismutase (SOD) and malondialdehyde (MDA), glutathione peroxidase (GSH-PX) and nitric oxide (NO) kits were obtained from Nanjing Jian Cheng Institute of Biological Engineering (Nanjing, China). Tumour necrosis factor-α (TNF-α) and interleukin-6 Enzyme-Linked Immuno Sorbent Assay (ELISA) kits were purchased from Suzhou Calvin Biotechnology Co., Ltd. (Suzhou, China).
Moloney Murine Leukemia Virus (M-MLV) Reverse Transcriptase, SYBR Premix Ex Taq II and DL2000 DNA Marker were supplied by TaKaRa (Dalian, China). Primary antibodies of ACC1 and LPL were purchased from Abcam (Cambridge, Britain). Antibody of GAPDH and HRP-conjugated goat anti rabbit IgG and goat anti mice were procured from Tiandeyue (Beijing) Biotechnology Co., Ltd. (Beijing, China).

The genomic DNAs (gDNAs) of bacteria used in the reference assays were extracted by chemical methods using a TIANamp Bacteria DNA Kit (Tiangen Biotech, Beijing, China). The concentrations of the purified gDNAs were calculated using a Nanodrop1000 spectrophotometer (ThermoFisher Scientific, USA) and diluted with water to desired concentrations. The 2 × LAMP mastermix was provided by CapitalBio Corporation (Beijing, China), and bovine serum albumin (BSA, final concentration of 3 mg mL⁻¹) was added to the mastermix (total volume of 60 μL) to decrease the non-specific adsorption of enzymes^{40 (link)}. Calcein, manganese chloride, and BSA were purchased from Sigma-Aldrich (Shanghai, China). The DL 2000 DNA marker was purchased from TaKaRa (Dalian, China), and GeneGreen dye was purchased from Tiangen Biotech (Beijing, China). The gel and chip images were obtained and processed using a gel imager (C150, Azure Biosystems, USA). All LAMP primer pairs were synthesized by Invitrogen (Beijing, China), and the sequences are listed in Supplementary Table S1. Each LAMP reaction required four to six primers named F3, B3, FIP, BIP, LF, and LB (LF and LB are not necessary), and the final concentration of each primer in our experiments was 0.3, 0.3, 2.4, 2.4, 1, and 1 μM, respectively.

The qRT-PCR products of cDNA and gDNA were investigated using 2% agarose gel electrophoresis with TAE running buffer. DNA was separated by electrophoresis at 100 V for 30 min. The DNA marker was DL2000 DNA Marker (2000-100 bp) (Takara). The bands were examined by UV irradiation.

Monoclonal antibodies against Bcl-2, Bax, P53, P21, Mcl-1, and GAPDH were purchased from Santa Cruz (Santa Cruz, CA, USA). Antibodies against CytC, Fas, FasL, and TRAIL were purchased from Abcam (Cambridge, MA, USA). Mouse anti-γ-H2AX antibody was from Millipore (Billerica, MA, USA). DMEM medium and FBS were purchased from Gibco BRL (Grand Island, NY, USA). FITC annexin V Apopotosis Detection Kit was from BD Biosciences (San Jose, CA, USA). Caspase-Glo 3/7, 8, and 9 assay kits and Plasmid extraction kit were purchased from Promega (Madison, WI, USA). N-acetyl-L-cysteine (NAC) and Reactive Oxygen Species Assay Kit were purchased from Sigma-Aldrich (St. Louis, MO, USA). Lipofectamine^TM 2000, Dichloro-dihydro-fluorescein diacetate (DCFH-DA), MitoSOX^TM Red, MitoTracker Deep Red FM, MitoTracker Green FM and Hoechst 33342 were from Invitrogen (Carlsbad, CA, USA). Trypan Blue Staining Cell Viability Assay Kit was from Beyotime Institute of Biotechnology (Jiangsu, China). Tetramethylrhodamine methyl ester (TMRM) was from Life Technologies (Carlsbad, CA, USA). Z-LEHD-FMK, Z-IETD-FMK, Z-DEVD-FMK were from Calbiochem (San Diego, CA, USA). Colorimetric TUNEL Apoptosis Assay Kit was from Beyotime (Shanghai, China). Mitochondria/Cytosol Fractionation Kit was from BioVision (San Francisco, CA, USA). DL-2000 DNA Marker was from TaKaRa Biotechnology (Dalian, China).

Normal Nutrient Agar, MacConkey Agar, and Eosin Methylene Blue Agar (EMB) were purchased from Beijing Aoboxing Biotechnology Co., Ltd. (Beijing, China).; DL 2000 DNA Marker was purchased from TAKARA; 2×Taq PCR Mix was purchased from Beijing Yuanchen Technology Co., Ltd. (Beijing, China). Gentamicin (GEN, 10 μg), spectinomycin (SPT, 100 μg), tetracycline (TE, 30 μg), ampicillin (AMP, 10 μg), ceftazidime (TAZ, 30 μg), ofloxacin (FOX, 5 μg), cotrimoxazole (SXT, 1.25/23.75 μg), and other drug-sensitive Discs were purchased from Hangzhou Binhe Microbiological Reagent Co., Ltd. (Hangzhou, China).; amoxicillin/clavulanic acid (AMC, 20/10 μg), florfenicol (FFC, 30 μg), meropenem (MEM, 10 μg), enrofloxacin (ENR, 5 μg), and sulfisoxazole (SIZ, 250/300 μg) were purchased from Hangzhou Microbial Reagent Co., Ltd. (Hangzhou, China).; apramycin (APR, 30 μg), polymyxin (CT, 10 μg), ceftiofur (EFT, 30 μg), and mequindox (SPI, 30 μg) were purchased from Yinuokang Technology Development Co., Ltd. (Tianjin, China). Escherichia coli quality control strain (25922, ATCC, USA) was purchased from Beina Chuanglian Biotechnology Co., Ltd. (Beijing, China).