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Fig. 4. (A) Reproducibility of the single cell RT-PCR protocol. Ventral marginal
zone (VMZ) and dorsal marginal zone (DMZ) explants were dissected and single
cells dissociated in calcium/magnesium-free medium. Single cells are picked
and lysed, then the lysate was split into A and B samples. These samples were
then subjected to RT-PCR in parallel. cDNA from each reaction was dot blotted
onto a nylon filter and probed with a specific marker. (B) Examples of dot
blots. cDNA from each single cell probed for the markers indicated (ODC,
Mix.1, Gsc, Xbra). (C) Sensitivity of single cell RT-PCR protocol. Single
cell lysates were spiked with 
1, 10, 50, 100, 500, 1000 and 5000
transcripts of polyadenylated eGFP RNA and subjected to RT-PCR, dot blotted
and probed for GFP. In two separate RT-PCR experiments, 10 transcripts of
GFP were detected. No signal above background was detected in the no GFP RNA
sample. (C) A dot blot where samples from separate RT-PCR reactions were
analyzed together.
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