Tiangen reverse transcriptase kit

Total RNA was extracted from different tissues of lettuce or Arabidopsis using Quick RNA isolation Kit (Waryoung, China). The RNA of wild type and LsFT-RNAi lines were extracted from the fourth leaves at 25 days. A TIANGEN reverse transcriptase kit (Tiangen) was used to synthesize cDNA. An ABI PRISM 7500 Real-Time PCR System (Applied Biosystems, Carlsbad, CA, United States) was used for quantitative real-time PCR (qRT-PCR) experiments. Three biological replicates and three technical replicates (3 × 3) were performed for each gene. Lettuce 18srRNA (Gene Bank accession number HM047292.1) and Arabidopsis ACTIN2 (Gene Bank accession number AT3G18780.2) genes were used as internal controls to normalize expression data. The gene-specific primers are listed in Supplementary Table S1.

Total RNA was extracted from mature leaves or flower buds using a Quick-RNA isolation Kit (Waryoung, Beijing, China). A TIANGEN reverse transcriptase kit (Tiangen Biotech, Beijing, China) was used to synthesize cDNA. Sequence information of LsFT (Lsat_1_v5_gn2_17881.1), LsAP1 (Lsat_1_v5_gn3_97021.1), LsAP3 (Lsat_1_v5_gn3_75340.1) and LsLFY (Lsat_1_v5_gn4_ 84380) were obtained by homologous alignment in the lettuce website³. AP1, AP3, and LFY were the downstream genes of FT (Jack et al., 1992 (link); Wagner et al., 1999 (link); Ferrandiz et al., 2000 (link)). Coding sequences (CDS) of LsAP1, LsAP3 and LsLFY were cloned from the shoot apex of bolting-sensitive S39 line and LsFT was choned from the mature leaves of S39 using gene specific primers (Supplementary Table S1).