alias: itsE5ITS2SS2P
title: ITS2: A. sarmentosa
design: DNA was extracted from the entire recruit for the small sized seven months old samples, and from a ~20 mg fragments collected from the coral branch tip for the adult and two years old juvenile samples. The extraction was conducted using a salting-out method with bead beating and additional enzymatic digestion steps with Lysozyme and Proteinase K. A tissue-free extraction was also conducted as a contamination control. Symbiodiniaceae ITS2 primers itsD [5?-TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGGTGAATTGCAGAACTCCGTG-3'] and Its2rev2 [5?-GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAGCCTCCGCTTACTTATATGCTT-3?] (Pochon, Pawlowski, Zaninetti, & Rowan, 2001) with Illumina adapter sequences (underlined) added at the 5-prime ends were used to amplify the partial 5.8S, entire ITS2 and partial 28S rDNA genes. Demultiplexed FASTQ files of all samples were processed in the same run and read quality was visualized using FastQC (Andrews, 2018). Conservative quality trimming was conducted using Trimmomatic and sequence reads were joined using PandaSeq (Masella, Bartram, Truszkowski, Brown, & Neufeld, 2012). Note that the files provided here have already been through conservative quality trimming and the forward and reserved reads have been joined.
identifiers.namespace: SUB5841729
identifiers.id: itsE5ITS2SS2P