title: Pacbio of  Tiarella polyphylla
design: 1. Reverse transcribe qualified RNA using SMARTer PCR cDNA Synthesis Kit to reverse transcribe RNA into cDNA; 2. PCR amplification using KAPA HiFi PCR Kits: A. PCR Optimization: optimize the PCR cycle number and select an optimal cycle number. B. Size Selection: PCR amplification is carried out according to the optimized cycle number, and the amplified products are mixed according to 0.5-6K gradient for fragment sorting.C, Large Scale PCR: The sorted fragments were amplified by Large Scale PCR to obtain sufficient total cDNA.3. SMRTbell Template Prep Kit 1.0 was used to construct SMRTbell library. Damage repair, end repair; Stem circular sequencing adaptors were connected at both ends of the cDNA fragment, and exonuclease was used to remove the failed fragment.