Carbon fixation by photosynthetic mats along a temperature gradient in a Tengchong hot spring
Zhang, Yidi (2020), Carbon fixation by photosynthetic mats along a temperature gradient in a Tengchong hot spring, Dryad, Dataset, https://doi.org/10.5061/dryad.r4xgxd28j
File of 13C-Raw data.xlsx
To measure the δ13C values of organic carbon, all samples were thawed at room temperature and centrifuged at 10,000 rpm for 15 minutes to collect the solid (biomass and sediment). The samples were acidified with 1 mM HCl overnight to remove inorganic carbon. The acidified samples were washed with deionized water to neutralize pH, and then dried at 50 ºC overnight. The dried solid samples were homogenized and weighted into sealed tin cups. δ13C values were measured twice for each sample with a Picarro iTOC-CRDS isotope analyzer.
File of DRCB.fasta
About 15 ml mat and surface sediment samples were collected for microbial community analysis. Samples were collected from sites DRCB-1, DRCB-2, DRCB-3, DRCB-4, DRCB-5 with sterilized spoons, and transferred into 50 ml polypropylene tubes. Genomic DNA was extracted from 0.5 wet sample using the FastDNA SPIN Kit (MP Biomedical, Solon, OH, USA) according to manufacturer’s protocol (Hou et al., 2013). PCR amplification was conducted with modified primers 515F (5’-GTG YCA GCM GCC GCG GTA A-3’) and 806R (5’-GGA CTA CHV GGG TWT CTA AT-3’) designed to be universal for bacteria and archaea (Caporaso et al., 2011; Caporaso et al., 2012). In order to identify individual samples from the reads, unique 8-bp barcodes were added at the 5′-end of both the forward and reverse primers. PCR products were purified following previously published literature (Hou et al., 2013). All PCR products were pooled together with equal molar amounts. Illumina MiSeq sequencing was performed with the MiSeq Reagent Kit v2 2× 250 bp.
Sequence demultiplexing and quality control were performed with Cutadapt 1.9.1 (Martin, 2011). The sequences that had an average quality score of lower than 27 were removed from subsequent analysis. Operational taxonomic unit (OTU) cluster at 97% sequence identity was determined by using UCLUST algorithm (Edgar, 2010). The first sequence from each OTU was picked as a representative, and taxonomy was assigned to each representative using the ribosome database project (RDP) classifier algorithm (Wang et al., 2007). Sequences that could not be classified with this algorithm were manually searched against the NCBI BLAST database using BlastN to find highly similar hits.
National Natural Science Foundation of China (NSFC), Award: No. 91851116