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| Hey all, i try to establish a sybr q pcr wich detects two different genes. i´ve already cloned my genes and a standard dilution was done. Followed by optimization of MgCl2 and pimer concentration. Due to the reason that i try to quantify aqueous bacteria via this two genes i designed and IC which runs with the same pcr conditions. But my question is: I calculate via the cp of my standard dilution my concentration of desired genes. Ok, but if there is an influence like inhibition due to the seawater i can detect this via my IC. So how would you calculate the inhibition of the desired genes in the sample in relation to the IC. For example: Standard 1E2 concentration with Cp 24; sample 1 5E1 concentration with Cp 27. IC in a.d. Cp 24; IC in environmetal sample Cp 30. This means a delay of 6 Cycles of the IC. So there is an influence visible but how should i implement this delay in final sample concentration calculation? Thanks for your help |
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| internal , quantification , standard |
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