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DNA-free CRISPR-Cas9 genome engineering in zebrafish
Amanda Haas, Alex J. Blasky*, Rytis Prekeris*, John A. Schiel, Melissa L. Kelley, and Anja van Brabant Smith | Dharmacon, now part of GE Healthcare, 2650 Crescent Drive, Suite 100, Lafayette, CO 80026, USA *University of Colorado - Anschutz Medical Campus, Department of Cell & Developmental Biology, Denver, CO, USA

Poster describing the advantages of a DNA-free gene editing system and the application of this system in zebrafish.

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Analysis of Terpenes Using Gas Chromatography with Vacuum Ultraviolet
Changling Qiu, Jonathan Smuts, Phillip Walsh, and Kevin A. Schug

The VUV absorption spectra for different terpenes were distinctive and differentiable. GC-VUV demonstrated the capabilities for qualitative and quantitative analysis of terpenes in turpertine mixtures. Chromatographic coeluting signals can be deconvolved by the VUV data analysis software.

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600 base reads on the Ion S5™ Next-Generation Sequencing System enables accurate HLA typing of 96 samples on one 530™ chip
Peter B. Vander Horn, Cisilya Duncan, Jamsheed Ghadiri, Amneet Gulati, Diana Jeon, April Jung, Mindy Landes, Tommie Lincecum, Geoffrey Lowman, Vadim Mozhayskiy, Linus Ong, Xinzhan Peng, Maryam Shenasa, Prasanna Thwar

We have demonstrated that by combining improvements in templating and sequencing biochemistry we are able to sequence templates longer than 600 bases with high accuracy on an Ion S5 530 chip.
These improvements open the S5 use space to include haplotyping applications that require longer reads. As a demonstration of that, we accurately typed 96 HLA samples on one 530 chip.

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Assessing Diversity in Cassava through the Application of Metabolomics
Margit Drapala, Elisabete Carvalhoa, Laura Perez-Fonsa, Elliott Pricea, L. Augusto Becerra Lopez-Lavalleb, Paul D. Frasera

In the present study metabolomic platforms have been established for Cassava and used to assess the biodiversity present in Cassava germplasm collections and elucidate underlying biochemical mechanisms associated with traits of interest.

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Metabolic response to everolimus in patient-derived xenografts of triple negative breast cancer
Leslie R. Euceda1, Deborah K. Hill1,2, Endre Stokke1, Elisabetta Marangoni3, Tone F. Bathen1, Siver A. Moestue1,2

Everolimus treatment metabolic effects on TNBC PDX models were investigated using MR spectroscopy. PLS-DA successfully discriminated treated from controls and PDX expressing INPP4B and not. LMM revealed significant differences in 4/17 metabolites and two metabolite ratios between treated and controls.

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Design considerations for highly specific and efficient synthetic crRNA molecules
Anja van Brabant Smith, Emily M. Anderson, Shawn McClelland, Elena Maksimova, Tyler Reed, Steve Lenger, Žaklina Strezoska, Hidevaldo Machado Dharmacon, part of GE Healthcare, 2650 Crescent Drive, Suite #100, Lafayette, CO 80026, US

An overview of our rational design algorithm for picking highly functional crRNA sequences in combination with comprehensive specificity analysis.

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Picking the best CRISPR-Cas9 targets for functional gene knockout: a machine learning algorithm based on both specificity and functionality
Shawn McClelland, Emily M. Anderson, Žaklina Strezoska, Elena Maksimova, Annaleen Vermeulen, Steve Lenger, Tyler Reed, and Anja van Brabant Smith Dharmacon, now part of GE Healthcare, 2650 Crescent Drive, Suite #100, Lafayette, CO 80026, US

The CRISPR-Cas9 system has the potential to significantly advance basic and applied research.

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Scaffold design, function and over-expression of lentiviral-based microRNAs
Angela Schoolmeesters, Melissa L. Kelley, Annaleen Vermeulen, Anja Smith, *Mayya Shveygert, *Xin Zhou, *Robert Blelloch Dharmacon, now part of GE Healthcare, 2650 Crescent Drive, Suite #100, Lafayette, CO 80026, USA

Here we describe the strategy for scaffold design, the importance of an optimal promoter, and demonstrate gene target down-regulation from the over-expression of lentiviral microRNA mimics.

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Homology-directed repair with Dharmacon™ Edit-R™ CRISPR-Cas9 and single-stranded DNA oligos
John A. Schiel, Eldon T. Chou, Maren Mayer, Emily M. Anderson , and Anja van Brabant Smith | Dharmacon, now part of GE Healthcare, 2650 Crescent Drive, Suite #100, Lafayette, CO 80026, US

Here we demonstrate how to perform lipid based transfections for homology directed repair using DharmaFECT Duo, CRISPR-Cas9 reagents and, synthetic DNA donor oligos.

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