Evaluation of methods for amplification of picogram amounts of total RNA for whole genome expression profiling

BMC Genomics

Table 1 aRNA and cDNA yields obtained after amplification using universal RNA, positive and/or negative controls.

Protocol	Sample	Starting amount	Amount of amplified material
Ambion	Universal RNA	500 pg	88.1 μg
			73.9 μg
	Universal RNA	250 pg	65.1 μg
			50.5 μg
	Negative control	0 pg	2.5 μg
Arcturus	Universal RNA	500 pg	34.3 μg
			15.3 μg
	Universal RNA	250 pg	45.6 μg
			16.2 μg
	Negative control	0 pg	1.9 μg
	Positive control RNA	500 pg	24.3 μg
			19.7 μg
Epicentre	Universal RNA	500 pg	55.6 μg
			6.3 μg
	Universal RNA	250 pg	9.4 μg
	Negative control	0 pg	1.9 μg
	Positive control RNA	500 pg	5.1 μg
			7.4 μg
Nugen	Universal RNA	500 pg	7.6 μg
			7.4 μg
	Universal RNA	250 pg	6.9 μg
			8.3 μg
	Negative control	0 pg	1.8 μg

For each protocol, one RNA amplification was performed from 250 pg and one from 500 pg of human universal RNA by two operators in two independent laboratories. Arcturus and Epicentre kits contain an internal positive control RNA. These RNA were systematically included when performing experiments with those chemistries. Negative control experiments (amplification without total RNA) were also performed to evaluate template-independent nucleic acid synthesis.

ISSN: 1471-2164