Arabidopsis

Now that the Arabidopsis thaliana genome has been sequenced, plant biologists are turning their efforts to genome annotation. In a 21 March report on the Sciencexpress website, Motoaki Seki and colleagues from the RIKEN Institute in Japan, describe a large-scale analysis of full-length Arabidopsis cDNA libraries (Sciencexpress 2002 10.1126/science.1071006).Seki et al. isolated over 150,000 RIKEN Arabidopsis Full-Length (RAFL) cDNA clones, sequenced the ends and clustered them into almost 15,000

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Now that the Arabidopsis thaliana genome has been sequenced, plant biologists are turning their efforts to genome annotation. In a 21 March report on the Sciencexpress website, Motoaki Seki and colleagues from the RIKEN Institute in Japan, describe a large-scale analysis of full-length Arabidopsis cDNA libraries (Sciencexpress 2002 10.1126/science.1071006).

Seki et al. isolated over 150,000 RIKEN Arabidopsis Full-Length (RAFL) cDNA clones, sequenced the ends and clustered them into almost 15,000 non-redundant groups. To generate the clones they constructed 19 cDNA libraries from plants grown under various conditions of stress, hormone and light. To optimize library construction they used the biotinylated CAP trapper method, using trehalose-thermoactivated reverse transcriptase combined with a single-strand linker ligation step, and normalization and subtraction procedures. They also created a database of promoter sequences upstream of RAFL clones. Amongst the clones were 837 that had been missed by previous annotation efforts; this project therefore makes a significant ...

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