Dictyostelium discoideum

Developmental cDNA Library

(Vegetative Stage )

Library construction

1. mRNA were prepared from AX4 cells at vegetative growth phase.
2. Oligo-dT primed cDNA synthesis with NotI and SalI linkers on ends.
3. After size fractionation with CHROMA SPIN-400 column (CLONTECH), the cDNA was ligated into pBluescript II KS-.
4. The ligated plasmid DNA was transformed into E. coli DH5 alpha cells.
5. 10752 clones were randomly picked and replicated on gridded filters.
6. After eliminating frequently appeared genes by probing the filters with selected genes, the remaining 8148 clones were subjected to sequencing.
   

Clone and sequence information

• Nucleotide sequences (csm.fasta)
• Clone information
• Search programs (FASTA, BLAST)