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To support our approach we developed a methodology based on three main steps (i.e. design, simulation and experimental validation) which can be easily carried out using the computing tool BioNetCAD.
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Using the "compute" tool, a homozygosity score was given for each site by taking the square of the difference of the frequencies of the alternate and reference alleles.
Based on the combined gene list, we first converted the gene IDs to protein IDs (UniProt Accession Number) to meet the requirement of the online tool, then calculated the protein isoelectric point (pI) and molecular weight (Mw) for all the corresponding proteins using the Compute pI/Mw tool provided by Expert Protein Analysis System (ExPASy, http://web.expasy.org/compute_pi/).org/compute_pi/
The theoretical isoelectric point and molecular weight were calculated by using the Compute pI/Mw tool of ExPASy.
The molecular weight and pI were predicted by using the Compute pI/Mw tool on the ExPASy Bioformatics Resources Portal (http://web.expasy.org/compute_pi/).org/compute_pi/
The theoretical molecular weight and isoelectric point were calculated using the compute pI/MW tool (http://www.expasy.org/tools/pi_tool.html/).html/
Theoretical average molecular weights were calculated from the expected protein sequences using the Compute Mw tool on the ExPASy Server.
The values of WM and pI were computed using the Compute pI/Mw tool (http://www.expasy.org/tools/pi_tool.html).html
The MW and pI calculations were performed using the Compute pI/Mw tool [ 24].
Predicted molecular weights for the opsins were calculated using the "Compute pI/Mw tool" on the ExPASy Proteomics Server [ 46].
The pI and molecular weight were estimated using the Compute pI/Mw tool from ExPASy (http://web.expasy.org/compute_pi).org/compute_pi
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