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Figure 4 | Plant Methods

Figure 4

From: Normalization using ploidy and genomic DNA copy number allows absolute quantification of transcripts, proteins and metabolites in cells

Figure 4

Quantification of RBC-L protein and genomic copy numbers in protein extracts from A. thaliana. (A) Standard curve indicating Ct as a function of the copy number of the cloned PCR product that was added to PCR reactions containing 1 μl of a 1:40 dilution of wild-type protein extract. Shown in the graph is an example of observed Ct values as a function of exogenously added DNA molecules. Each reaction contained 1 μl of a 1:40 dilution of wild-type protein extract and an arbitrary amount of external plasmid (p) harboring DNA fragments amplified by PCR using the primers T7F6-3-F and T7F6-3-R (see additional file 1). (B) SDS-PAGE analysis of RBC-L accumulation in A. thaliana. The indicated number of purified recombinant RBC-L molecules (lanes a to e) and the indicated amount of wild-type protein extract sample (lanes f to i) was loaded, and the gel was stained with CBB. The arrow indicates RBC-L migration. (C) Standard curve indicating the number of recombinant RBC-L molecules relative to the intensity of CBB staining.

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