Dictyostelium discoideum

Developmental cDNA Library

(First Finger Stage )

Library construction

1. mRNA preparation from AX4 cells at first finger stage (14 h-16h) of development
2. oligo-dT primed cDNA synthesis with NotI and SalI linkers on ends
3. Size fractionation to yield two sublibraries, Library S (formerly named as Library B) and Library L
4. Ligation into pBluescript II KS- (Library S) or pSPORT1 vector (Library L)
5. Transformation into E. coli DH5 alpha (Library S) or DL795 cells (Library L)
6. Picking up 9984 clones from Library S and 8448 clones from Library L
7. Preparation of gridded filters which carry the picked clones

Library specification:、

	Library S	Library L
Vector	pBluescript II KS-	pSPORT 1
Strain of E. coli	DH5 alpha	DL795
Size (cfu/ml)	6 x 10E6	2.5 x 10E6
Mean insert (kbp)	1	1.3
No. of clones picked	9984	8448

Caution!: SSA101 - SSA196 have older names SL-X001 - SL-X096

Clone and sequence information

We attempt to sequence 10,000 rondomely chosen clones from the libraries.
Sequences were read only once from 3'and 5'.

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