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Fig. 2. (A) The single cell RT-PCR protocol. Ventral marginal zone (VMZ), dorsal marginal zone (DMZ), vegetal and animal pole explants are dissected and single cells dissociated in calcium/magnesium-free medium. Single cells are picked and subjected to RT-PCR. cDNA from each single cell is run on an agarose gel to check integrity and amplification, then Southern blotted or dot blotted, and probed with a specific marker. (B) Examples of dot blots. cDNA from each single cell is dotted onto a nylon filter in the format of a 96-well microtitre plate and probed for the markers indicated (ODC, Xbra, XK70A, Sox17{alpha}, Xwnt8, Gsc). In this example, cells from stage 10 DMZ and VMZ explants are shown; column/row number corresponds to those shown for dorsal and ventral cells in Fig. 3.





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