DNA Isolation from Cells
Cell culture are important for research of disease and basic science. The ability to isolate high-quality DNA from a variety of cell cultures is important. The PoP team members have developed a protocol using Beckman Coulter reagents to isolate both DNA from cultured cell samples.
DNA extraction from Jurkart cells using GenFind V3
Data shown below resulted from isolated genomic DNA (gDNA) from 1.4 million Jurkart, an immortalized line of human T lymphocyte cells. DNA yield and purity was assessed by a NanoDrop (Thermo Fisher Scientific) [Table 1].
|Cell Number||Conc. (ng/µL)||Yield (µg)||A260/A280||A260/A230|
|Jurkat cells (Sample A)||1.4 x 106||323.8||13.0||1.98||2.16|
|Jurkat cells (Sample B)||1.4 x 106||319.9||12.8||1.97||2.15|
|Jurkat cells (Sample C)||1.4 x 106
|Average||1.4 x 106
The genomic integrity of the three replicates was assessed on an Agilent Genomic DNA Screen Tape (Agilent). The DIN scores, which represent the amount of gDNA degradation, were all above 8.4 indicating highly intact gDNA predominately greater than 48.5 kb (Figure 1). An example electropherogram of sample C can be seen in figure 2.
Beckman Coulter makes no warranties of any kind whatsoever express or implied, with respect to this protocol, including but not limited to warranties of fitness for a particular purpose or merchantability or that the protocol is non-infringing. All warranties are expressly disclaimed. Your use of the method is solely at your own risk, without recourse to Beckman Coulter. Not intended or validated or use in the diagnosis of disease or other conditions. This protocol is for demonstration only, and is not validated by Beckman Coulter.
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