Grant number: | 98/12250-0 |
Support Opportunities: | Genome Research Grants |
Start date: | December 01, 1998 |
End date: | May 31, 2004 |
Field of knowledge: | Biological Sciences - Biochemistry - Molecular Biology |
Principal Investigator: | Paulo Arruda |
Grantee: | Paulo Arruda |
Host Institution: | Centro de Biologia Molecular e Engenharia Genética (CBMEG). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil |
Abstract
The objective of this proposal is to construct cDNA libraries for The Sugarcane EST Project (SUCEST) and organize strategies to store and distribute the clones for sequencing. Sequencing will be performed by the Organization for Nucleotide Sequencing and Analysis (ONSA) according to the rules established in SUCEST (see accompanying text). Normalized cDNA libraries will be prepared from RNA extracted from sugarcane tissues (calli, root, stalk, etiolated leaves, flowers, developing seed, "in vitro" growing plantlets, etc) according to the methodologies established by Dr. Marcelo Bento Soares, Department of Pediatrics, College of Medicine, University of Iowa. The quality of the libraries, in terms of insert size and redundancy, will be checked by sequencing and PCR analysis. The libraries that passed the quality control will be ordered in microplates, replicated and distributed for sequencing by the ONSA network. A replica of each clone will be maintained by the DNA coordinator until the sequences are deposited in the database. The information from the database will then be used by the DNA coordinator to re-order the clones to be sent to the Storage and Distribution Coordinator. This strategy will be used to sequence up to 50,000 different sugarcane ESTs or when 200,000 readings are deposited in the database. Information on the overall project organization can be seen on the accompanying SUCEST text. (AU)
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