Single Specific Primer-PCR

For SSP-PCR, cDNA of de-etiolated or etiolated Arabidopsis seedlings was used as template, and divergent forward primer and divergent reverse primer were added for first round of PCR, respectively. These fragments were amplified by first round of PCR using the Pyrobest DNA polymerase (TaKaRa, Dalian, China) for 35 cycles in a 50 μl reaction with these primers (Supplementary Table 2). The reactions were amplified using the qRT-PCR mix using the “Quantitative real-time PCR” program on a Biorad thermocycler.