Primestar max premix 2 dna polymerase

The DNA manipulation procedure was performed as described by Green and Sambrook [27 ]. Genomic DNA was isolated using TIANamp Bacteria DNA kit (Tiangen Biotech, Beijing, China) according to the manufacturer’s instructions. Plasmids were prepared using the high-purity plasmid small quantity preparation kit (Tiangen Biotech, Beijing, China), following the manufacturer’s instructions. DNA fragments encoding styAB gene and mdh gene were amplified by PCR using PrimeSTAR Max Premix (2×) DNA polymerase (Takara Bio, Dalian, China) according to the manufacturer’s protocol, with the following cycle profile: 95 °C for 5 min, 40 cycles of 94 °C for 30 s, 53 °C for 1 min, 72 °C for 5 s, and a final extension at 72 °C. Primers used are shown in Table S1.

Indigo pigment standard products, tryptophan, N, N-dimethylformamide, indole and kanamycin were purchased from Sigma–Aldrich (St. Louis, MO, USA); PrimeSTAR Max Premix (2×) DNA polymerase, T4 DNA ligase, nucleic acid restriction quickCut enzyme HindIII, BamHI, MluI, XbaI, SfoI, MluI and molecular quality standard DNA Marker, 10,000 DL DNA Marker, were purchased from Takara Bio (Dalian, China). Other reagents are domestic market, pure analysis.