Two-fold differences are the detection limit for determining transgene copy numbers in plants by real-time PCR

BMC Biotechnology

Table 1 Copy number analysis for progeny of 13 homozygous lines (two plants for each line) with two different calibrators.

Construct	Plant	ΔCt (Ct FAM - Ct TET) ± SD^a	Copy number with A434-4-2 as calibrator	Copy number with A300-1-2 as calibrator	Copy number from Southern analysis
as-hpl	A188-4-1	0.42 ± 0.11	4.8	1.4	1
	A188-4-2	2.26 ± 1.77	1.4	0.4	1
	A247-3-1	0.28 ± 0.07	5.3	1.6	2
	A247-3-2	0.72 ± 0.10	3.9	1.2	2
	A337-2-1	0.89 ± 0.24	3.5	1.0	1
	A337-2-2	1.80 ± 0.44	1.9	0.5	1
	A422-4-1	1.76 ± 0.63	1.9	0.6	1
	A422-4-2	3.17 ± 0.34	0.7	0.2	1
	A434-4-1	2.69 ± 1.36	1.0	0.3	1
	A434-4-2	2.69 ± 0.95	1.0	0.3	1
as-aos	A160-5-1	0.69 ± 0.06	4.0	1.2	1
	A160-5-2	1.13 ± 0.32	3.0	0.9	1
	A160-7-1	1.10 ± 0.07	3.0	0.9	1
	A160-7-2	1.88 ± 0.14	1.8	0.5	1
	A266-2-1	2.36 ± 0.35	1.3	0.4	1
	A266-2-2	1.91 ± 0.25	1.7	0.5	1
	A340-1-1	1.32 ± 0.64	2.6	0.8	1
	A340-1-2	1.02 ± 0.25	3.2	0.9	1
as-lox	A211-1-2-1	2.62 ± 0.27	1.2	0.3	1
	A211-1-2-2	2.46 ± 0.36	1.1	0.3	1
	A248-2-1	-0.17 ± 0.22	7.2	2.1	2
	A248-2-2	0.74 ± 0.73	3.9	1.1	2
	A300-1-1	1.42 ± 0.69	2.4	0.7	1
	A300-1-2	0.93 ± 0.75	3.4	1.0	1
	A363-1-1	-0.06 ± 0.39	6.7	2.0	2
	A363-1-2	-0.32 ± 0.16	8.1	2.4	2
controls	Wildtype 12	12.04 ± 0.81	0	0	-
	A340-5-1^b	8.51 ± 0.10	0	0	-
	A340-5-2^b	9.02 ± 0.26	0	0	-

^a average of three measurements ^b transformed line without T-DNA (T-DNA segregated out)

ISSN: 1472-6750