PRKAA2 mRNA levels were measured using quantitative real-time PCR (qPCR). Total RNA was extracted using an RNA extraction reagent (Takara, 9108, Japan), and 2X Super SYBR Green qPCR Master Mix (ES Science, Guangzhou, China) was used to detect PRKAA2 levels. The qPCR primer sequences ...
采用PCV2体外感染小鼠脾淋巴细胞，在培养液中加入终浓度为100、200和400mg/L 的SSP，分别培养8、12和24h 后收取样品，荧光定量PCR 技术测定细胞内炎性因子的mRNA 表达水平。结果显示，当小鼠脾淋巴细胞培养至24h 时，PCV2感染对照组的NF-κB、MCP-1、CXCL10和STAT1的表达水平显著高于细胞对照组（P <0.05）；...
The following RT–PCR kits were tested to determine the best performing assay: (1) OneStep RT–PCR Kit (QIAGEN); (2) TaqPath 1-Step Multiplex Master Mix (Thermo Fisher Scientific); (3) One-Step PrimeScript RT–PCR Kit (Takara Bio); (4) GoTaq Probe qPCR Kit (Promega Corporation); (...
PCR products were subcloned into the pTA2 vector (TOYOBO) and sequenced. Real-time RT–PCR analysis. Total RNA was isolated from cells using TRIzol reagent (Invitrogen) and 1 mg of total RNA was reverse transcribed to obtain first- strand cDNA using an RNA-PCR kit (Takara) following the ...
针对KRT2基因设计特异性siRNA-1/2，转染A549细胞后采用细胞活力检测试剂盒检测下调KRT2蛋白表达后对A549细胞活力影响，进一步利用荧光定量PCR 方法检测干扰效率，筛选最佳siRNA ，结果显示，与对照siRNA 相比，KRT2特异性siRNA 处理不影响A549细胞活力，且siRNA-2能更有效降低KRT2的表达。以A/WSN/1933（H1N1）株病毒...
2.7. Quantitative real-time PCR (qRT-PCR) analysis RNA in the cell culture supernatant was extracted with Libex 96 (qEx-DNA/RNA, T183X) and used for reverse transcription by PrimerScript RT Master Mix (Takara, Lot:AN82328A). qRT-PCR was performed on a CFX Connect (Bio-rad) using iTa...
The 20 μL PCR reaction mixture was generated according to manufacturer instructions (Takara Ex Taq®, Takara, Dalian, China). The cis-elements in the JrGRAS2 promoter were categorized into different groups after the analysis using the online programs PLACE [58] and PLANTCARE [59]. The ...
For capillary sequencing with a 3130xl Genetic Analyzer (Thermo Fisher Scientific), PCR products prepared from 10 ng of template cDNA were used to amplify CD74-NRG2α and FGFR2-MBIP by GoTaq G2 Hot Start Master Mix Green (Promega, Madison, WI), in accordance with the manufacturer’s ins...
Macropinocytosis is essential for myeloid cells to survey their environment and for growth of RAS-transformed cancer cells. Several growth factors and inflammatory stimuli are known to induce macropinocytosis, but its endogenous inhibitors have remained
As the COVID-19 infection continues to ravage the world, the advent of an efficient as well as the economization of the existing RT-PCR based detection assay essentially can become a blessing in these testing times and significantly help in the management of the pandemic. This study demonstrat...