Zebrafish EST Vector

Clones are streaked onto LB agar containing ampicillin (50µg/ml). To use: re-streak the clone onto the same agar type, in such a way as to obtain single colonies. Incubate at 37°C overnight. Clones can be grown in LB broth + ampicillin, and glycerol added after growth to a concentration of 25% for long-term storage at -70°C.

cDNA was oligo dT primed from 8 ug of total RNA isolated from FACS sorted GFP positive myoblast cells taken from 3-9 somite stage Tg(alpha-actin:GFP) embryos. SalI_NotI cut cDNA was then ligated into pSPORT1 with SalI at the 5' end and NotI at the 3' end.