He has over 35 years of general management, consulting, and program management experience implementing knowledge processes, knowledge management platforms, and content enrichment solutions.
This is particularly important given the fact that different target enrichment solutions have its own pros and cons.
We show that the new protocol combines the simplicity of PCR amplicon sequencing with the accuracy of molecular barcodes, can provide deep coverage for a very large region, and will be a useful addition to existing target enrichment solutions.
Haemophilus influenzae were grown on chocolate agar and seeded into Brain heart infusion broth containing 10 ml/L IsoVitaleX enrichment solution (BD Biosciences), 5% fetal bovine serum (Invitrogen), and 25 mg/L hemin (Sigma).
Last ,100 mL of Middlebrook OADC enrichment solution (BD, Franklin Lakes, NJ, USA) was added aseptically to the medium after cooling to 50 55°C.
TGE may be performed in solution (solution-based targeted genomic enrichment, solution-based TGE, [ 5]), or on an array platform (solid-phase targeted genomic enrichment, solid-phase TGE, [ 6]).
Symbiodinium minutum, originally provided by Dr. Mary Alice Coffroth, of the University of New York, Buffalo, was cultured in autoclaved, artificial seawater containing 1X Guillard's (F/2) marine-water enrichment solution (Sigma-Aldrich: G0154), plus three antibiotics, ampicillin (100 μg/mL), kanamycin (50 μg/mL), and streptomycin (50 μg/mL) [ 18].
The sporophytes (conchocelis) of this pure line were directly developed from a single somatic cell that was enzymatically isolated from a farmed thallus and then cultured in bubbling sterilized seawater with Provasoli's enrichment solution medium (PES) under 20 μmol photons · m 2 s–1 at 24 ± 1 °C and a 12 12 light:dark (L D) photoperiod.
Genomic DNA was captured using the Agilent enrichment solution method (SureSelect Human All Exon Kit Version 2, Agilent) with the Agilent bank of biotinylated oligonucleotide probes (Human All Exon v2 - 50 Mb, Agilent), followed by a high-throughput sequencing of the 75 bases at each end on an Illumina HiSeq 2000 (Gnirke et al, 2009).
Next, RNA fragments larger than 150 nt are precipitated with 10 μl of enrichment solution (25% PEG 8000 (polyethylene glycol), 2.5 M sodium acetate in RNase-free water) on ice for 30 min and centrifuged at 16,000 × g for 20 min at 4°C.
Cells were grown in sterile F/2 media with sea salts (32 g/L, Sigma Aldrich), 40 mM Tris HCl (pH 8) and Guillard's (F/2) marine water enrichment solution (Sigma Aldrich) in Erlenmeyer flasks with 100 μmol photons m−2 s−1 illumination and 100 rpm agitation at 22 ± 1 °C in a growth chamber.
