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1) Retrieval of chromosomal locus matched the EST sequence by Blast search. EST sequence (mRNA). AAAAAAAAA. GATC. M. genomic DNA. 2) Search for the nearest Pst I site from MboI site of EST. P. M. 3) Design of the primers from Pst I site to fit defined conditions. P.
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1) Retrieval of chromosomal locus matched the EST sequence by Blast search EST sequence (mRNA) AAAAAAAAA GATC M genomic DNA 2) Search for the nearest Pst I site from MboI site of EST P M 3) Design of the primers from Pst I site to fit defined conditions P 4) Final selection of primer by Blast Search P