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SRX4404544: NSL_031_20160606_HLSU
1 ILLUMINA (Illumina MiSeq) run: 77 spots, 38,654 bases, 107,337b downloads

Design: Large subunit haptophyte-specific fusion primers (HAP_LSU_F: GGURUCGGAGARGGUGAGAAUCC and LHapto20R_bis: TCAGACTCCTTGGTCCGTGTTTCT) containing Illumina-specific adaptors with 6 unique inline barcodes for the forward primer and 8 indices for the reverse primer were used. Triplicate PCR reactions were carried out for each genomic DNA sample with a 33 cocktail of 1.0 U Platinum High-Fidelity DNA Polymerase (Life Technologies, Carlsbad, CA): with 1X Hi-Fidelity buffer, 200 dNTP mix, 1.5 mM MgSO4, and 0.2 of each primer. 5-20 ng of template DNA were added to each PCR and included a no-template control for the run. PCR conditions were as follows: 2 min at 94, 30 s at 94, 30 s at 55, 3 min 68, with repetition from step 2 for 35 cycles followed by 5 min at 68. Resulting PCR tripiclate reactions were pooled and products were verified using a Caliper LabChip GX. Amplicons were cleaned with Agencourt AMPure XP magnetic beads and quantified with Quant-iT PicoGreen dsDNA Assay Kit. The amplicons were equimolarly pooled and the size was verified using the Agilent Bioanalyzer DNA 1000 chip. In addition, the concentration of the amplicon pool was checked with qPCR. The pool of amplicon libraries was diluted to 13 pM prior to clustering, and PhiX was added as a diversity control for the MiSeq sequencing run.
Submitted by: Marine Biological Laboratory
Study: Global Group I Haptophytes - Large Subunit
show Abstracthide Abstract
The purpose of this project is to characterize the diversity of Group I Isochrysidales in Northern Hemisphere lakes, despite the conservation of the distinct biomarker signatures produced by these organisms.
Sample: Lake E1, Alaska
SAMN09475417 • SRS3558955 • All experiments • All runs
Library:
Name: NSL_031_20160606_HLSU
Instrument: Illumina MiSeq
Strategy: AMPLICON
Source: METAGENOMIC
Selection: PCR
Layout: PAIRED
Runs: 1 run, 77 spots, 38,654 bases, 107,337b
Run# of Spots# of BasesSizePublished
SRR75369727738,654107,337b2020-06-01

ID:
5986506

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