HglT fragments of heterocytous cyanobacteria

Assembled hglT gene fragments obtained via PCR and direct sequencing using primer set 2 (Fw1 mix B + Rv1, Supplementary table 2) on the heterocytous cyanobacterial cultures listed in Table 2, Supplementary Table 8 and shown in Figure 5 (sequences in blue) in Pérez Gallego et al. 2023 

Nucleotide sequences were obtained via Sanger sequencing and were processed using Geneious Prime (v 2023.0.4). Sequences were trimmed using an error probability limit of 0.01. Potential heterozygous bases in single reads were identified using a 50% peak similarity cutoff, peak detection height was set at 10%. When available, the consensus sequence was obtained by aligning forward and reverse reads with Geneious assembler using the highest sensitivity settings. When appropriate, sequences belonging to the pCR™4-TOPO™ vector (Invitrogen, Carlsbad, CA, USA) were identified and removed.

To generate the phylogenetic trees sequences were aligned using MAFFT (v7.407) with L-INS-i iterative refinement method (Katoh and Standley, 2013) and poorly aligned regions were removed using trimAl (Capella-Gutiérrez et al., 2009). Phylogenetic trees were built using IQ-tree (v1.6.7) and its in-built nucleotide substitution model finder (Kalyaanamoorthy et al., 2017), using 1000 replicates to perform SH-like approximate likelihood ratio test (SH-aLRT) (Guindon et al., 2010) and 1000 bootstrap replicates.