The cDNA library was constructed from mRNA isolated from differentiating somatic embryos cultured on MSM6AC. The library was prepared using the Stratagene pBluescript II SK (+) library construction kit. Complementary DNA was synthesized from mRNA using a primer consisting of a poly(dT) sequence with an Xho I restriction site. Eco RI adaptors were ligated to the blunt-ended cDNA fragments followed by Xho I digestion. The cDNA fragments were directionally cloned into the Eco RI-Xho I restriction site of the pBluescript vector. The ligated cDNA fragments were transformed into E. coli ElectroMax DH10B host cells. Tissue culture and library construction were performed by Françoise Thibaud-Nissen and Anu Khana (Lila Vodkin lab, University of Illinois).
Transformation efficiency: 1.5 x 106
Percent white colonies: 64%
Average insert size of white colonies: 910 bp
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