Improved Ligation Specificity with Chemically Modified Ligation Components
Poster Oct 04, 2012
Sabrina Shore, Alexandre Lebedev, Elena Hidalgo Ashrafi, Gerald Zon, Natasha Paul, Richard Hogrefe
Ligases are gaining utility in molecular biology applications, such as nucleotide sequence detection, single nucleotide polymorphism (SNP) detection, protein detection and “next generation” sequencing by ligation. With the increased demand for DNA ligases in the field of biotechnology, comes increased demand for ligation fidelity. Described approaches to improved ligation fidelity include ligases from different biological sources, point mutations of key amino acid residues within the ligase, modified reaction conditions and addition of crowding reagents, such as PEG. Although most approaches to improved ligation fidelity have focused on the ligase itself, further improvements are needed and may be attainable by a different approach. Herein a strategy to improve the discrimination between matched and mismatched targets is described which employs chemical modification to the nucleic acid components of the reaction, such as the donor probe, the acceptor probe and the ATP cofactor. The results demonstrate that chemically modified components increase the stringency of DNA ligase-mediated nucleic acid detection, providing a unique approach for SNP genotyping.
Complete “Sample-to-Result” Highly Automated NGS and qPCR Workflow for Clinical DiagnosticsPoster
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In summary, ISG15 expression levels are crucial for trophoblast morphology and function (migration/invasion). By blocking trophoblast invasion, reduced ISG15 levels could contribute to impaired spiral artery transformation that reduces utero-placental blood flow in preeclampsia. Thus, agents inducing ISG15 expression are likely to be therapeutic in preeclampsia.
An Emerging Phenotype of Partial RAG 1/2 Deficiency Among Young Children with Autoimmunity and Viral InfectionsPoster
We describe the natural history of a cohort of 12 patients with confirmed partial RAG1/2 mutations and autoimmunity at a young age. We were seeking the link between viral infections and autoimmunity and tested candidate biomarkers that may reflect the underlying RAG1/2 protein deficiency.READ MORE