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PCR products were bi-directionally sequenced on an Applied Biosystems 3730 DNA Analyzer and analyzed with SoftGenetic's Mutation Surveyor software.
Sequence differences to the NCBI reference sequence were identified via manual inspection of aligned electropherograms assisted by the Mutation Surveyor software package (SoftGenetics, State College, PA).
Chromatograms were analyzed for methylation quantification using the Mutation Surveyor Software (SOFTGENETICS®) with default parameters.
Data were analysed by visual inspection of electropherograms and using Mutation Surveyor software (SoftGenetics , Inc.
The sequence graphs were analysed manually and with Mutation Surveyor software (SoftGenetics, State College, PA, USA).
Sequences were analysed with Mutation Surveyor software (Softgenetics, State College, PA, USA).
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Sequencing electropherograms were analyzed by visual inspection of electropherograms and by Mutation Surveyor 2.03 Software (SoftGenetics).
The deletion junction sequences of all samples were further analyzed by PCR amplification across the deletion junction, followed by sequence analysis using the BigDye Terminator Cycle Sequencing kit (version 3.1) and the ABI3730XL automated DNA sequencer with Mutation Surveyor V3.20 software (Softgenetics, State College, CA).
Sequences were screened for variants using Mutation Surveyor v2.61 software (SoftGenetics, State College, PA, USA).
The sequences were analysed using ABI PRISM® 3100 GeneticAnalyzer/HITACHI (Applied Biosystems) and the presence of mutations were performed using the Mutation Surveyor (Softgenetics) software.
The cells were imaged daily with an Olympus IX71 microscope with the OASIS automation control system and Surveyor imaging software version 5.5.5.26 (Objective Imaging Ltd., Cambridge, UK).
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