Kod plus neo dna polymerase kit

Total RNA was extracted using Trizol (Tiangen, Beijing, China) from the trifoliolate leaves of ZGDD seedlings sampled on day 9 after commencing LD treatment; cDNA was synthesized with Superscript II reverse transcriptase (TransGen Biotech, Beijing, China) and used as a template for further GmFT1a cDNA cloning. Amplification was performed using PCR reactions with a KOD‐plus‐Neo DNA polymerase kit (Toyobo, Tokyo, Japan).

For GmFT1a cloning, 125 soybean varieties covering all known (14) MGs were collected (Table S4). Genomic DNA was extracted from these varieties. GmFT1a were amplified with a KOD‐plus‐Neo DNA polymerase kit (Toyobo) using the primers shown in Table S3. The PCR products were directly Sanger sequenced by the Tsingke Biological Technology Co. (Beijing, China). The resulting sequences were mapped to the soybean reference genome (v.Wm82.a2.v1) using BWA (v.0.7.10‐r789, using the ‘mem’ subcommand) with default settings, and GmFT1a polymorphisms were analyzed using Samtools (v.1.4.1, using the ‘mpileup’ subcommand) (http://samtools.sourceforge.net/) and bcftools v.1.4.1, using the ‘call’ subcommand) (https://github.com/samtools/bcftools).