2.3.2. 50
RACE
Total RNA of leaf tissue (1 mg) was used to synthesize the first
strand cDNA according to the manufacturer’s instructions of
the SMARTer RACE cDNA Amplification Kit User Manual
(Clontech Laboratories, Inc., TAKARA). The gene-specific
primers were designed and synthesized according to the
above sequenced 30 ends (Table 1). Then touchdown PCR
was performed using the following program: 5 cycles of 30 s
at 94 C and 90 s at 72 C, 5 cycles of 30 s at 94 C, 30 s at
70 C and 1 min at 72 C, 30 cycles of 30 s at 94 C, 30 s at
65 C and 120 s at 72 C (for RrNHX1). Conditions of the
other gene amplification are also listed in Table 1.