For mRNA detection, tissues collected were snap frozen and stored in liquid nitrogen until analysis. Approximately 20 – 100 mg of frozen breast tumor tissue was crushed in liquid nitrogen. RNA was extracted using commercial kits (Qiagen), RNA integrity was assessed on the Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA), cDNA was synthesized from 1 mg of total RNA using Invitrogen kits (Invitrogen Corp.).
Invitrogen kits
Manufactured by Thermo Fisher Scientific
Sourced in United States, China
Invitrogen kits are a range of laboratory products manufactured by Thermo Fisher Scientific. These kits are designed to provide researchers with tools for various applications, such as DNA and RNA manipulation, protein expression and purification, and cell culture. The core function of these kits is to enable researchers to perform specific experiments and analyses in a streamlined and efficient manner.
Automatically generated - may contain errors
3 protocols using invitrogen kits
1
mRNA Detection in Breast Tumor Tissue
2
Quantitative Analysis of RNA Expression
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The total RNA was extracted from homogenized liver tissues and L02 cells using TRIZOL (Invitrogen, Carlsbad, CA, United States). Both reverse transcription and quantitative PCR (RT-qPCR) were performed using the Invitrogen kits (Invitrogen, Shanghai, China) on the ABI Prism 7900 Sequence Detection System (Applied Biosystems, Alameda, CA, United States). Primers for tumor necrosis factor alpha (TNF-α), interleukin (IL)-6, IL-8, and FOXO3 were synthesized using Invitrogen (Invitrogen, Shanghai, China). For miR-122-5p expression analysis, TaqMan-based miRNA RT-PCR was used. The primer sequences used are listed in Table 1 . The mRNA and miRNA abundance were calculated using the 2–ΔΔCt method and normalized to β-actin and U6, respectively.
3
Cervical Brush DNA Extraction
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DNA extraction from the cervical brushes was performed using commercial Invitrogen ® kits following the manufacturer's recommendations, which included a previous step of proteinase K digestion. The extracted DNA was quantitated using a spectrophotometer Nanodrop 2000 ® (ThermoScientific, USA) .
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