Diversity and Biogeography of Bacteria in the Surface Sediments of the Western Arctic Ocean
PCR amplification of the 16S rRNA gene V6 fragments was performed with universal bacterial primers 967F (5'-CNACGCGAAGAACCTTANC-3') and 1046R (5'-CGACAGCCATGCANCACCT-3'). A set of ten nucleotide (nt) barcodes was designed and added to the 5’ end of 967F for the multiplexing of samples in the Solexa pair-end (PE) sequencing runs. After sequencing, a Perl script was used to separate the reads from each sample according to their barcode sequence in either of sequencing file, F or R(fastq format). The reads were overlapped to assemble the final V6 tag sequences with the global alignment using the Needleman-Wunsch (NW) algorithm. And then the assembled sequences were subjected to quality control and chimera screening. Finally, we got the clean sequences for the subsequent analyses.