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A combinatorial library of the original Anticalin was generated by error-prone PCR, subjected to E. coli cell surface display, and applied to repeated cycles of cell sorting after incubation with the fluorescently labelled target protein under competition with the unlabelled extracellular domain of CTLA-4.
It consists in the simultaneous amplification of several fluorescently labelled target amplicons (BRCA1 and BRCA2 exons) and reference sequences.
The first is based on the analysis of arrays which have been co-hybridised with three differently labelled target samples, with n = 5 arrays.
A reference design was used in which approximately 30 pmol of Cy3 labelled target RNA was hybridised against the common oligonucleotide reference (representing 30 pmol of Cy5).
The addition of Alexa 594 as a dye-label for an additional – third – target sample works within the framework of more commonplace Cy5/Cy3 labelled target sample combinations.
The eVOC ontology is a controlled vocabulary used to describe the sample source of cDNA and SAGE libraries and labelled target cDNAs for microarray experiments.
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As previously advised by N'Guyen [ 36], we first determined the amount of labelled target to be used for each hybridisation so that no additional signal appeared but the intensity of the positive signals increased when the amount of target did (50, 125, 250 and 500 ng for aRNA or cDNA labelled targets; data not shown).
The boosting process requires some amount of labeled target data.
Magnetite cationic liposomes (MCLs) were used to label target cells.
Finally, the target learner is built from the labeled and pseudo labeled target data.
For this scenario, there is an abundance of labeled source data and limited labeled target data.
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