Statistics based algorithm for exact quantitation of [biotechnology] seed traits without reference material

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“…#15 ( ) 25.93 0.06 t C = ± is the first after the median, which has Ct value that differs from #10 outside the error. Self-reference calibration through dilutions of aliquot #20 showed true linearity 2 and the number of target seeds (GTS 40-3-2 beans) in the premedian aliquot is n1≈6.98=7.…”

“…3000 balls-(16 red+14 blue+2970 black) emulating the mixture of A2704-12 and A5547-127, and (150 red+2850 black) emulating GTS 40-3-2. Mixture of 3000 balls was put into an opaque bag, thoroughly mixed, then random ball was taken out "blindly" by hand (like when playing lotto) and put into one of 20 numbered (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20) glasses. Then the balls left in the bag were mixed again.…”

“…The common and globally approved method of quantitative analysis of biotechnology traits/genetically modified organisms (GMO) is PCR in real time mode when the value of threshold cycle (C t ) of tested sample is compared with C t values obtained through sequential dilution of the corresponding certified (standard) reference material (CRM) presented in the form of linear regression in semilogarithmic co-ordinates C t vs log[concentration, %]. 1 There are recent developments on using digital PCR with direct DNA copy counting, 2 it also requires CRM though. However, certified reference materials may be quite expensive and not easily available, if available at all.…”