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10.1371/journal.pone.0252507 http://scihub22266oqcxt.onion/10.1371/journal.pone.0252507 34061896!8168896!34061896     free     free     free Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       PLoS+One 2021 ; 16 (6): e0252507 Nephropedia Template TP {{tp|p=34061896|t=2021. Producing molecular biology reagents without purification |pdf=|usr=}}{{34061896}} gab.com Text Producing molecular biology reagents without purification JO: PLoS One http://www.kidney.de/pget.php?&tw=1&pmid=34061896 #FOAvip We recently developed 'cellular' reagents-lyophilized bacteria overexpressing proteins of interest-that can replace commercial pure enzymes in typical diagnostic and molecular biology reactions. To make cellular reagent technology widely accessible and amenable to local production with minimal instrumentation, we now report a significantly simplified method for preparing cellular reagents that requires only a common bacterial incubator to grow and subsequently dry enzyme-expressing bacteria at 37 degrees C with the aid of inexpensive chemical desiccants. We demonstrate application of such dried cellular reagents in common molecular and synthetic biology processes, such as PCR, qPCR, reverse transcription, isothermal amplification, and Golden Gate DNA assembly, in building easy-to-use testing kits, and in rapid reagent production for meeting extraordinary diagnostic demands such as those being faced in the ongoing SARS-CoV-2 pandemic. Furthermore, we demonstrate feasibility of local production by successfully implementing this minimized procedure and preparing cellular reagents in several countries, including the United Kingdom, Cameroon, and Ghana. Our results demonstrate possibilities for readily scalable local and distributed reagent production, and further instantiate the opportunities available via synthetic biology in general. Twit Text FOAVip Producing molecular biology reagents without purification ????PLoS One http://www.kidney.de/pget.php?&tw=1&pmid=34061896 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=34061896 #FOAvip English Wikipedia <ref>{{Cite pmid|34061896}}</ref> Producing molecular biology reagents without purification #MMPMID34061896 Bhadra S; Nguyen V; Torres JA; Kar S; Fadanka S; Gandini C; Akligoh H; Paik I; Maranhao AC; Molloy J; Ellington AD PLoS One 2021[]; 16 (6): e0252507 PMID34061896show ga We recently developed 'cellular' reagents-lyophilized bacteria overexpressing proteins of interest-that can replace commercial pure enzymes in typical diagnostic and molecular biology reactions. To make cellular reagent technology widely accessible and amenable to local production with minimal instrumentation, we now report a significantly simplified method for preparing cellular reagents that requires only a common bacterial incubator to grow and subsequently dry enzyme-expressing bacteria at 37 degrees C with the aid of inexpensive chemical desiccants. We demonstrate application of such dried cellular reagents in common molecular and synthetic biology processes, such as PCR, qPCR, reverse transcription, isothermal amplification, and Golden Gate DNA assembly, in building easy-to-use testing kits, and in rapid reagent production for meeting extraordinary diagnostic demands such as those being faced in the ongoing SARS-CoV-2 pandemic. Furthermore, we demonstrate feasibility of local production by successfully implementing this minimized procedure and preparing cellular reagents in several countries, including the United Kingdom, Cameroon, and Ghana. Our results demonstrate possibilities for readily scalable local and distributed reagent production, and further instantiate the opportunities available via synthetic biology in general. |COVID-19 Testing/methods/*standards[MESH] |COVID-19/*diagnosis/*epidemiology/virology[MESH] |Cameroon/epidemiology[MESH] |Diagnostic Tests, Routine/*standards[MESH] |Escherichia coli/genetics/metabolism[MESH] |Gene Expression[MESH] |Geobacillus stearothermophilus/genetics/metabolism[MESH] |Ghana/epidemiology[MESH] |Humans[MESH] |Indicators and Reagents/chemistry/metabolism/*standards/supply & distribution[MESH] |Molecular Diagnostic Techniques[MESH] |Plasmids/chemistry/metabolism[MESH] |Real-Time Polymerase Chain Reaction/methods/*standards[MESH] |Recombinant Proteins/biosynthesis/genetics[MESH] |SARS-CoV-2/*genetics[MESH] |Synthetic Biology/methods[MESH] |Transformation, Bacterial[MESH]Producing molecular biology reagents without purification (epmc).pdf DeepDyve Pubget Overpricing