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.
Applications of chemically modified synthetic guide RNA for CRISPR-Cas9 genome editingPoster
Our results indicate that MS modifications are required for experiments with co-electroporation of Cas9 mRNA and synthetic gRNA, yet have no impact on editing efficiency when delivered with lipid-based transfection reagents.READ MORE
Automating Mass Spectrometry-Based Quantitative Glycomics using Tandem Mass Tag (TMT) Reagents with SimGlycanPoster
One of the emerging trends in glycomics research is the innovation related to accurate MS based quantitative analysis of glycans.READ MORE
Highly Accurate HCV Genotyping by Targeted Next Generation SequencingPoster
The recent fast advancement of next generation sequencing (NGS) technologies allowing for unprecedented speed and accuracy in analyzing viral genomes are opening new ways to further improve diagnostic genotyping of HCV.
Comments | 0 ADD COMMENT
Protein Discovered that Protects Against Fatty Liver
News Jan 18, 2017