Introduction

To study the function of a certain gene, a relatively common approach is to construct cell knockdown/overexpression models, and then investigate the changes at the transcriptome level as well as the genes, pathways and phenotypes regulated by it. The commonly used methods for constructing knockdown/overexpression cell models are: transient transfection of plasmids/shRNA/siRNA and lentiviral infection to prepare stable cell models.

However, different cells require different transfection conditions and methods. For different genes of interest, should knockdown or overexpression be chosen? How to conduct accurate quality control to ensure the reliability, authenticity, high quality and reproducibility of the subsequent sequencing data and phenotypic experimental results? These will become the key factors for accurately constructing cell knockdown/overexpression models.

advantages

✔ Our company has an experience library covering over a hundred kinds of cell transfection operations and projects, and can tailor the optimal transfection plan for the cell model you desire.
✔ With strict quality control at every step, we can ensure the reliability, authenticity, high quality and reproducibility of the subsequent sequencing data and phenotypic experimental results.
✔ We provide data analysis, graphing, and English materials and methods, as well as detailed raw data, to safeguard your article publication.
✔ Examples of published articles.
✔ Our company has experience in over a hundred kinds of cell culture and transfection, covering various disease types, as shown in the table on the right (including but not limited to these).

Workflow

Delivery Contents:
A. Raw data of all experimental results.
B. Experimental reports (including detailed experimental procedures and the manufacturer and catalog number information of the main instruments and reagents).
C. English materials and methods.

Demo Data

Taking the knockdown of human umbilical vein endothelial cells (HUVECs) as an example.

Lentiviral transfection
Detection of RNA integrity - agarose gel electrophoresis / RIN (RNA Integrity Number) / RQN (RNA Quality Number) value detection: RQN / RIN ≥ 7; The three bands in the gel electrophoresis are clear, and the brightness of the 28S band is twice that of the 18S band.
RNA quantification: A. Purity: The ratio of A260/A280 is between 1.8 and 2.2; the ratio of A260/A230 is between 1 and 2. B. Total amount and concentration: The total amount is ≥ 2 μg, and the concentration is ≥ 100 ng/μl.
Detection by RT-qPCR: The results of QPCR showed that at the mRNA level, the knockdown was over 70%, and the T-test was significant.
WB detection: At the protein level of the target gene, the knockdown was successful, and the T-test was significant.