Our volume that contain 0.1ug of our sample plasmid DNA were 2.4154ul and 0.7189ul respectively.
How should I do the comparison, please
Dear Mathew,
I would set both plasmids to same concentration. As you had the lowest, use your concentration and dilute your colleagues sample to your concentration, then compare the two side by side with controles.
Also, I liked the answer above too, run them on a gel and look.
Best Wishes!
Hi Mathew,
139 ng/ul is a decent yield using a mini prep kit. If the elution volume is the same for both samples then 41 ng/ul is not great; but atleast you were able to extract the plasmid.
Small variances from culture volume, lysis time, passing the supernatant through the column twice and heating the elution buffer before adding to the resin can influence the yield.
If you want just comparison your colleague got better quantity 139 ng/ul of DNA than you 41 ng/ul.
Did you check it on gel !!!
Or if you want to proceed for further for e.g. sequencing your colleague need to dilute that but you can directly use without dilution.
What exactly you want to do with that!!
You mentioned the following values 2.4154ul and 0.7189ul. Even quantity wise also it is very less. How did you measure so exactly the volumes!!!
Dear Mathew,
I would set both plasmids to same concentration. As you had the lowest, use your concentration and dilute your colleagues sample to your concentration, then compare the two side by side with controles.
Also, I liked the answer above too, run them on a gel and look.
Best Wishes!
ug are masses, no concentrations. There is not much to compare, you just can say I have less than she/he. If you rather mean ug/ul, then check if you have the same volumes where you dissolved your DNA (if not, calculate how much DNA you both actually have). If you have started from the *same* culture, you may state that your colleague has a "better" yield, but there may be RNA contamination, so just figures form an OD260 won't say anything without further analysis. If you started from different cultures, you just may state that she/he had a higher yield than you. Without replicates, you can't do any statistics and can't make any statement about the culture or the quality of the overall prep itself. It just was a single experiment.
I think you should compare the elution volume,run on a gel, or do seq.
Helal F Hetta Wolfgang Schechinger Charles D Anderson Jetty Ramadevi Praveesh Valissery Amanda Koryl Please find the complete table of the group and lecturer said I should discuss my result in comparison with other students results. How can I go about it? I hope its clear enough now. Thank you in anticipation
Lots of data. You can do ranking, award prizes for it, calculate means and medians and percentiles and stuff. But what is the purpose? Usually, people are happy when there is enough for the downstream application (normally: yes) and when - more important - it's pure enough and is really the plasmid you want (to this end, we sequence it or do restriction digests or PCRs).
The major conclusion IMHO is that if you have a lot of people doing an experiment, you'll get a lot of different results. What do you think might be the reason for this variation if you look at all steps involved in the prep?
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