RNase If -treated quantitative PCR for dsRNA quantitation of RNAi trait in genetically modified crops

BMC Biotechnology

Table 3 RNase I_f-qPCR assay to analyze distinct ratios of dsRNA and ssRNA mixtures

Mixture	dsRNA to ssRNA ratio	Means of dsRNA RTL	Means of ssRNA RTL	Expected dsRNA to ssRNA RTL ratio	Means of measured dsRNA to ssRNA RTL ratio^a
1	1:5	31.27	148.52	0.20	0.21
2	2:7	25.49	75.07	0.29	0,34
3	3:8	21.44	45.67	0.38	0.47
4	1:2	21.92	38.30	0.50	0.57
5	1:1.5	17.67	23.29	0.67	0.76
6	10:1	567.75	56.41	10.00	10.06
7	5:1	115.43	23.21	5.00	4.97
8	7:2	85.94	22.33	3.50	3.85
9	1:1	48.00	37.15	1.00	1.29

RNase I_f -qPCR assay was performed to quantify the mixed synthetic dsRNA and ssRNA with distinct ratios indicated above, and the RTL of dsRNA and ssRNA was analyzed, respectively. Each mixtures were analyzed at least two technical replicates and the calculated mean ratios were presented. The dsRNA and ssRNA RTL were measured by R95 RTL and 70 RTL, respectively
^aChi-square analysis showed no significant difference between the expected and measured ratios

ISSN: 1472-6750