The cDNAs were extracted by boiling the gel slices, ethanol precipitating the cDNAs, and redissolving them in water. Those cells which contained successfully ligated clones were grown up and their plasmid DNA was extracted and precipitated. The sequences obtained were then compared with the known gene sequences contained in the GenBank database. 3 Top is uncut SA6 plasmid, lane 3 Bottom is uncut 4SA6 plasmid. Differential Display gel using 4 cell lines.
Molecular size is indicated by the numbers on the left of the gel.
These include cDNA bands representing mRNAs that were produced in one or more of the four cell lines, as well as those produced in different amounts in the different cell lines as is apparent by differences in the intensity of the bands produced. The differential expression of mRNAs between various NB cell lines allowed us to continue our investigation into the genetic make-up of NBs as well as specific genetic differences between the individual NB cell lines. We successfully isolated 38 cDNA bands from polyacrylamide gets. These bands included cDNAs from each of the four cell lines. PCR amplification of these cDNA clones resulted in eleven differentially expressed cDNAs which we were able to clone into phagemid vectors.