http://www.scientistsolutions.com/t13189-dna+copy+number+and+calculating+_+gmo.html Life Science Discussion en-us sci7feed@sci7.nojunkorherepleasespam.com hourly 1 2005-05-05T12:00+00:00 http://www.scientistsolutions.com/t13189-dna+copy+number+and+calculating+_+gmo.html Best regards,
Monique]]> Mon, 16 Nov 2009 00:56:26 GMT http://www.scientistsolutions.com/t13189-dna+copy+number+and+calculating+_+gmo.html Hi Monique,
Unfortunately what you are trying to do is highly dependent on the assay you have and the RNA/DNA standards you are using. If you do not have a good assay, or very well characterized template standards, then it is going to be very tricky to get the assay to work successfully.
Unfortunately I am not aware of an assay you could do for this as most of these assays are designed specifically for the given GMO being tested.
Sincerely,
I...]]> Sun, 15 Nov 2009 15:50:47 GMT http://www.scientistsolutions.com/t13189-dna+copy+number+and+calculating+_+gmo.html I have already tried to work with % gm (mixing) but I don't know what is going wrong.
Purpose of my experiment is to get % gmo from unknown samples.
After DNA isolation, samples are going into RT PCR. I use ISO21570 protocol for quantification.
My standards (when I use CRM %# don't show logic Ct values, for example I have higher Ct for higher concentrations.#).
So I have a lot of problems...If you have some experiences with good protocols, Ill
a...]]> Sat, 14 Nov 2009 05:45:18 GMT http://www.scientistsolutions.com/t13189-dna+copy+number+and+calculating+_+gmo.html Hi Monique,
What you are trying to do is a little tricky, but here is what I suggest. You already have qPCR assays for your GMO modification and for an internal standard (reference gene). Now what you need is a DNA sample that you know does not have any GMO in it (reference sample), and if possible a second reference sample that is 100% GMO. I would even go as far as mixing your 0% GMP sample with your 100% GMO sample in different quantities so that instead of running a standard...]]> Thu, 12 Nov 2009 10:27:48 GMT http://www.scientistsolutions.com/t13189-dna+copy+number+and+calculating+_+gmo.html I am new in q real time pcr and I have problem.
I don't know to calculate a % of gmo based on copy numbers.
I do relative quantification, for example rapeseed, I have one standard curve (based on copy numbers 300000,30000,3000,300,30) for reference genes and another for gmo modification.
I do not know what to do with my quantities.
Or If I have value in copies how to convert it in % of gmo?
]]> Thu, 12 Nov 2009 08:03:47 GMT