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The first step toward developing the high throughput platform was to curate all known hordeins sequences from EMBL Nucleotide Sequence Database (EMBL), DNA Data Bank of Japan (DDBJ), GenBank at the NCBI (GenBank) and HarvEST databases.
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The Probesets were annotated by searching against the Arabidopsis genome using the tool provided in HarvEST database (http://harvest-web.org).
The annotations of the Barley1 GeneChip were updated by the HarvEST database [ 52].
In order to approach this large genome, several projects have generated significant numbers of expressed sequence tags (ESTs) (ca. 500 000) (see HarvEST database http://harvest.ucr.edu/).ucr.edu/
For this analysis we used the HarvEST database which contains over 50,000 unigenes and searched for miRNA precursors corresponding to smRNA sequences in our database.
Since there are additional markers on chromosome 3H mapped in other germplasm [ 34] (see also HarvEST database [ 3]), more 3H BAC clones could be sequenced.
We also compared the new citrus ESTs with the ESTs and unigenes already available in the Citrus HarvEST database, which contains 229,570 ESTs.
Finally, the new citrus ESTs collection added almost 2000 new genes not previously represented in the Citrus HarvEST database, which already contains 229,570 ESTs.
The HarvEST database (http://harvest.ucr.edu/) (17), dedicated to several crop species, including T. aestivum and Hordeum vulgare, provides access to curated EST assemblies, comparative analysis tools and links to orthologues in related model plant species.
Throughout this study, a custom microarray SCRI-Hv35-44k-v1 (Agilent design 020599) representing approx. 42 000 barley unigene sequences from the public HarvEST database (assembly 35; http://www.harvest-web.org/) was used (ArrayExpress accession A-MEXP-1728 A-MEXP-1728 A-MEXP-1728
To link the expressed signatures to known genes from citrus, the unigene dataset from TIGR gene index database and Harvest database were combined to create a reference gene dataset.
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