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Abrahams, G. L.

Publications and source records attributed to Abrahams, G. L..

2 recordsLinked to original sources

AG3D: A low-cost educational 3D printable toolkit for agarose gel electrophoresis

Access to desktop additive manufacturing is growing and the argument could be made for 3D printers to be standard laboratory equipment. The power of the printers lies in the democratisation of scientific equipment. Traditional agarose gel electrophoresis forms a cornerstone of molecular biology research, teaching and learning. Reliable electrophoresis is dependent on a number of factors which include standardized commercial equipment with respect to casting trays, combs, horizontal tanks and power supplies. The aforementioned systems come at a high initial cost; this is before factoring in the costs of standard electrophoresis grade agarose and associated reagent pricing. Here, we present a basic method for the additive manufacturing of a simple 3D printable agarose gel electrophoresis (AGE) unit with a built-in gel casting tray for standard size microscope slide-based AGE; named AG3D. The system presented was validated using different standard agarose-buffers (Tris Acetate EDTA and Tris Borate EDTA) and commercially available base-pair ladders. We provide a comparison between the AG3D and a commercial AGE system in respect to resolving power of the electrophoresis unit and discuss the overall reduction in cost afforded by the AG3D electrophoresis toolkit. The method presented has the potential for application in low resource educational environments by: O_LISignificantly lowering costs through the reduction of reagents (agarose, buffers etc). C_LIO_LIAllow for the use of low sample volumes. C_LIO_LIProviding an open-source toolkit for modification whether for research or teaching & learning C_LI Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=64 SRC="FIGDIR/small/533785v1_ufig1.gif" ALT="Figure 1"> View larger version (16K): org.highwire.dtl.DTLVardef@135b120org.highwire.dtl.DTLVardef@1ffca3eorg.highwire.dtl.DTLVardef@36ce4corg.highwire.dtl.DTLVardef@21dfb0_HPS_FORMAT_FIGEXP M_FIG C_FIG Specifications table O_TBL View this table: org.highwire.dtl.DTLVardef@1e4a30forg.highwire.dtl.DTLVardef@18d57c9org.highwire.dtl.DTLVardef@7e8b77org.highwire.dtl.DTLVardef@1f64979org.highwire.dtl.DTLVardef@51e36f_HPS_FORMAT_FIGEXP M_TBL C_TBL

scientific communication and education↗

DNA-dependent binding of nargenicin to DnaE1 inhibits replication in Mycobacterium tuberculosis

Natural products provide a rich source of potential antimicrobials for use in treating infectious diseases for which drug resistance has emerged. Foremost among these is tuberculosis. Assessment of the antimycobacterial activity of nargenicin, a natural product that targets the replicative DNA polymerase of Staphylococcus aureus, revealed that it is a bactericidal genotoxin that induces a DNA damage response in Mycobacterium tuberculosis (Mtb) and inhibits growth by blocking the replicative DNA polymerase, DnaE1. Cryo-electron microscopy revealed that binding of nargenicin to Mtb DnaE1 requires the DNA substrate such that nargenicin is wedged between the terminal base pair and the polymerase and occupies the position of both the incoming nucleotide and templating base. Comparative analysis across three bacterial species suggests that the activity of nargenicin is partly attributable to the DNA binding affinity of the replicative polymerase. This work has laid the foundation for target-led drug discovery efforts focused on Mtb DnaE1.

microbiology↗