DNA molecular probe and application in miRNA real-time quantitative PCR detection in tumor cells of green bean nuclease
A DNA molecule, mung bean nuclease technology, applied in the field of miRNA detection in tumor cells, can solve the problems of increasing experimental errors, many experimental steps, time-consuming detection errors, etc.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0048] 1) The experiment was divided into a normal cell control group (human bronchial epithelial HBE cells) and a tumor cell experimental group (human non-small cell lung cancer A549 cells). The culture and other experimental operating conditions of the two groups of cells were the same. It is expected that the theoretical length of the PCR amplification product is about 65bp.
[0049] 2) subculture human non-small cell lung cancer A549 cells and human bronchial epithelial HBE cells into 6-well cell culture plates, and routinely culture them in DMEM medium containing 10% calf serum (volume percentage) (95% humidity, 37°C , 5% CO 2 (volume percentage) until the cell growth reaches 80% confluency.
[0050] 3) The cell culture medium was discarded, and the remaining cell culture medium was washed away with PBS buffer, then Trizol reagent was added, and the total RNA of A549 cells and human bronchial epithelial HBE cells was extracted according to the standard total RNA extracti...
Embodiment 2
[0068] Other cases of tumor cell proliferation detection:
[0069] 1) The experiment was divided into normal cell control group (breast normal epithelial cells MCF-10A–parental, human embryonic kidney epithelial 293T cells) and tumor cell experimental group (human breast cancer MDA-MB-231 cells and human cervical cancer Hela cells), The culture and other experimental operating conditions of the two groups of cells were the same. It is expected that the theoretical length of the PCR amplification product is about 65bp.
[0070] 2) Subculture MCF-10A-parental, MDA-MB-231, 293T and Hela cells into 6-well cell culture plates respectively, and perform cell culture according to the method described in Example 1.
[0071] 3) The method described in Example 1 was used to set up positive controls and negative controls, and Beta-Actin was used as the sample internal reference.
[0072] 4) The method described in Example 1 was used to perform experimental operations such as RNA extract...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More