Is it necessary to rate 260 to 280 plasmid after digested to be 1.8 then do transformation Pichia pastoris by electroporation?
I use Phenol: chloroform: isoamyl alcohol, but I cant get suitable answer (260/280=1.8)
How can I improve it?
Its a range any ratio between 1.6-1.8 is good. Your downstream process should work.
Hi Karim,
The 260/280 ratio gives you an idea of the purity of your DNA on the ratio of DNA (260) to protein (280). You can get away with DNA that is not so pure for some applications, for other applications purity is critical. Do you have the ability to use a plasmid purification kit? If not, it might make sense to try to use new reagents to see if you might be able to get your ratio closer to 1.8.
I agree with the comments given above. Although 260/280 ratios are important indicators of sample purity and quality, the best indicator of DNA or RNA quality is functionality in the downstream application of interest. 1.8 is fine for me. You can go ahead with the next downstream step, i.e., transformation.
Excuse me, you mean I ignore 260/280 ratios for transformation Pichia.
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