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dna purity is 17? what does it mean?Moderator: BioTeam
7 posts • Page 1 of 1
dna purity is 17? what does it mean?Hi everyone,
I'm constructing a new vector and before starting ligation I measured dna purity of my gene. The dna purity that I measured is 17. More more than 1.8 Thanks for yor help
Re: dna purity is 17? what does it mean?Yes I'm sure that it is 17 not 1.7. The spectrophotometer calculates automaticly when I enter my dilution ratio. I used spectro lots of times and that is the first time ı see a value like this. But ı make dna cleaning and concentrating prosedure and forget my dna sample 5 or 7 minutes in the final step(elution step). Is this the matter I really don't undestand??
you're centrifuging it for 5 minutes?
Anyway, what was your concentration? Such out-of-mind values usually come up when you have very low concentration and thus very low absorbance. BTW with the 260/280 ratio higher than 2, the DNA is contaminated with some RNA, phenols or something like that. But I doubt anyone ever saw 17 http://www.biolib.cz/en/main/
Cis or trans? That's what matters.
Re: dna purity is 17? what does it mean?Interestingly my dna concentration is 157,07 mikrogram/ml
I also want to ask another question like this problem. Today I purified DNA from pellets. Then I read their purity and concentrations in spectrophotometer. The purities range between 1,8-1,9. Really good and concentrations are good also. Then I digest this samples with an enzyme and look if it digest, the digestion was perfect. I cut the bands that I want and the problem starts at this point: I made gel extraction and now the purities range between 3,5-5 . The concentrations are 5 mikrogram/ml. I'm so unhappy that you can't guess after the perfect purity and concentration from first step. Is it the agarose that wastes my samples? What will I do?
7 posts • Page 1 of 1
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