Standardization of DNA Residual Quantification Method of Vero Cell Rabies Vaccine for Human Use

Table 4: RT-PCR standardization for B-actin. Effect of the concentration of MgCl₂ and primers on Ct.

Template	[MgCl₂]	*[Primers]*	Ct
pDNA 1 x10⁷ Copies/µl	2mM	25	37,02
gDNA Vero 50ng/µl			38,66
NTC			39,95
pDNA 1 x10⁷ Copies/µl		40	22,27
gDNA Vero 50ng/µl			29,72
NTC			40
pDNA 1 x10⁷Copies/µl		50	17,29
gDNA Vero 50ng/µl			24,16
NTC			27,41*
pDNA 1 x10⁷ Copies/µl		75	16,46
gDNA Vero 50ng/µl			24,13
NTC			40
pDNA 1 x10⁷ Copies/µl	1.75mM	25	40
gDNA Vero 50ng/µl			40
NTC			40
pDNA 1 x10⁷Copies/µl		40	26,17
gDNA Vero 50ng/µl			35,82
NTC			40
pDNA 1 x10⁷ Copies/µl		50	40
gDNA Vero 50ng/µl			40
NTC			40
pDNA 1 x10⁷Copies/µl		75	21,57
gDNA Vero 50ng/µl			28
NTC			33,69
pDNA 1 x10⁷ copies/µl	1.5mM	25	40
gDNA Vero 50ng/µl			38,56
NTC			40
pDNA 1 x10⁷ copies/µl		40	35,9
gDNA Vero 50ng/µl			34,8
NTC			40
pDNA 1 x10⁷ copies/µl		50	31,07
gDNA Vero 50ng/µl			38,59
NTC			40
pDNA 1 x10⁷ copies/µl		75	35,54
gDNA Vero 50ng/µl			35,96
NTC			37,87

* The signal observed for NTC template correspond to primers dimers and was not observed in subsequent assays.