This primer library contains 96 sets of gene-specific primers that can be used for expression profiling of 96 genes, of which 93 are Arabidopsis pathogen-inducible genes and 3 are housekeeping genes (Appendix). It is designed for expression analysis of Arabidopsis defense response genes by quantitative RT-PCR (Q-RT-PCR). The pathogen-inducible genes are selected based on a search of an Arabidopsis database and microarray analysis (Appendix for the list of Primer names). The housekeeping genes, Actin-2, CBP (nuclear cap-binding protein) 20, and Ubiquitin, are selected based on their expression patterns.
- Packaged in a 96-well plate format.
- Each well contains a lyophilized set of forward and reverse primers for a specific gene.
- Each primer set allows for 200 RT-PCR reactions (20-25 µL reaction volume).
- Suitable for one-step or two-step RT-PCR.
- Works for both real-time and end-point time RT-PCR analysis.
The specificity and amplification efficiency of all primers have been validated using pathogen-infected Arabidopsis leaf samples and SYBR® Green Quantitative RT-PCR Kit (QR0100) (Figures 1 & 2).
Figure 1. Real-time quantification of a defense-response gene expression.
The expression level of the defense gene was highly induced in pathogen treated samples (32 folds of increase) compared to the untreated sample; whereas the internal control actin-2 gene has the same expression level (same CT value) in both samples.
Figure 2. Real-time RT-PCR amplification of all 96 selected genes in pathogen treated and untreated Arabidopsis leaves.
Many genes are induced in pathogen treated samples as indicated by reduced CT values. The CT for pathogen treated sample ranged from cycle 23 to 31, whereas the CT from untreated control ranged from cycle 27 to 33. The data was normalized against control gene amplifications (Table 1).
Table 1
Expression patterns upon Pseudomonas syringae (Pst or expressing avrRpm1) infection in Arabidopsis leaves at different time points.
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