Quantitative study of genomic DNA in HEK293 cell line by digital PCR
10.13200/j.cnki.cjb.004431
- VernacularTitle:数字PCR技术对HEK293细胞系基因组DNA的定量
- Author:
BI Hua
- Publication Type:Journal Article
- Keywords:
HEK293;
Digital PCR;
Host DNA;
DNA quantification
- From:
Chinese Journal of Biologicals
2025;28(02):190-196+203
- CountryChina
- Language:Chinese
-
Abstract:
Objective To perform quantirative study of HEK293 cell genomic DNA by microfluidic chip-based digital PCR technology, and to provide new ideas for more accurate gene quantification and genomic DNA related detection.Methods Firstly, the HEK293 cell DNA was obtained by Spin Column, identified and analyzed for the purity by agarose gel electrophoresis. Then, the genomic DNA of HEK293 cells was quantified by traditional ultraviolet spectrophotometry, Qubit fluorescence quantification and microfluidic chip-based digital PCR technology and statistically analyzed.Results The concentration of genomic DNA in HEK293 cells was detected as 100. 08 ng/μL by ultraviolet spectrophotometry, 93. 98 ng/μL by Qubit fluorescence quantification and the copy number was determined as 29 722. 81 copies/μL by digital PCR. Roughly calculated based on human single-copy genome of about 3. 3 pg, the converted copy numbers of spectrophotometry and Qubit fluorescence quantification were about 30 327 and 28 479 copies/μL, respectively. The deviation of the digital PCR detection value from the spectrophotometry detection value was only 2%, and the deviation from the Qubit detection value was only4%.Conclusion In this study, HEK293 genomic DNA was detected and compared by digital PCR, spectrophotometry and Qubit fluorescence, which provides a new detection method and idea for DNA quantification, and also provides a certain reference for the quantification of other nucleic acids.
