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. 1998 Aug 4;95(16):9620–9625. doi: 10.1073/pnas.95.16.9620

Figure 1.

Figure 1

Hybridization signals are quantitatively related to the amount of RNA template used for aRNA amplification. (A) Three λ DNA (i.e., 0.5-, 2.0-, and 6.6-kb HindIII fragments) in plasmid vector were spotted twice on each blot (1 μg of plasmid DNA on each spot). These three synthetic λ RNAs were added at four concentrations. Blots: a, 20 ng/μl; b, 4 ng/μl; c, 0.8 ng/μl; d, 0.16 ng/μl. Blots were probed with labeled aRNA (Left). After stripping, the same blots were reprobed with an oligonucleotide probe to determine the amount of plasmid DNA on each spot (Right). Because plasmid DNAs were not added in equal molar amounts, the molarity of plasmid DNA for the one with shorter insert should be higher (Right). (B) Normalized hybridization signal is linear to the amount of RNA template used for amplification. The amount of λ RNA is expressed as molar concentration.