Alkaline hydrolysis to remove potentially infectious viral RNA contaminants from DNA

Table 3 Effects of safety treatment on FMDV P1 cDNA sequencing applications

Sample ID	Heat	SCR	NaOH	HCl	TE/ H₂O	P1 Sequence
1- FMD P1 cDNA heated	x	0 μl	0 μl	0 μl	37 μl	+
2- FMD P1 cDNA + SCR heated	x	7 μl	0 μl	0 μl	30 μl	+
3- FMD P1 cDNA treated	x	0 μl	15 μl	15 μl	7 μl	+
4- FMD P1 cDNA + SCR treated	x	7 μl	15 μl	15 μl	0 μl	+
5- FMD P1 cDNA diluted in H₂O		0 μl	0 μl	0 μl	37 μl	+
6- FMD P1 cDNA diluted salt		0 μl	15 μl^a	15 μl^a	7 μl	+
7- FMD P1 cDNA control		0 μl	0 μl	0 μl	0 μl	+

Samples containing various combinations of FMDV cDNA, SCR, and/or treatment conditions. FMDV sequencing results of 35 μl FMDV cDNA spanning the P1 coding region of the genome were determined to verify cDNA integrity under different treatment conditions. NaOH and heat treatment incubation time used in this study was 60 min as optimized in previous studies. ^aA neutralized salt solution containing equal parts of NaOH and HCl was added instead of individual base and acid treatments to serve as a positive control. X indicates 65 °C for 60 min and 10 min boiling steps were included, and + indicates positive sequencing results

ISSN: 1743-422X