Optimized differential display and reamplification parameters for silver staining.
Differential display (DD) is a powerful instrument to detect differences in gene expression of malignant and normal cells. An optimized silver staining protocol was used to compare mRNA expression of keratinocytes and squamous cell carcinoma cells of the head and neck. Optimal concentration for upstream and downstream primer was 0.2 microM. Best primer concentrations for reamplification were between 40 and 60 pM. With this optimized protocol nearly 50 differentially expressed transcripts have been identified and differential display can be applied safer, easier, and faster, than by radioactive labeling.
Gottschlich
S
S
Department of Otorhinolaryngology, Head and Neck Surgery, Christian-Albrechts-University Kiel, Germany.
Goeroegh
T
T
Folz
B J
BJ
Lippert
B M
BM
Werner
J A
JA
D002294:Carcinoma, Squamous Cell / Q000235:genetics*; D018076:DNA, Complementary / Q000096:biosynthesis; D006258:Head and Neck Neoplasms / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger / Q000096:biosynthesis; D016622:Silver Staining; D014158:Transcription, Genetic / Q000235:genetics; D014407:Tumor Cells, Cultured
D018076:DNA, Complementary; D012333:RNA, Messenger