Representative cDNA libraries and their utility in gene expression profiling.
An increasing interest in gene expression profiles in human diseases has led to the use of microdissected tumors and biopsies in gene discovery approaches. Since many of these clinical samples yield extremely small amounts of RNA, reproducible methods are needed to amplify this RNA while maintaining the original message profile. Using the SMART cDNA Synthesis Method, we show that high-, medium- and low-abundance transcripts can be amplified in a representative fashion and that the resulting cDNA can also be used as a complex probe to confirm gene expression differences identified by other techniques.
Endege
W O
WO
Chiron Diagnostics, East Walpole, MA, USA.
Steinmann
K E
KE
Boardman
L A
LA
Thibodeau
S N
SN
Schlegel
R
R
D018076:DNA, Complementary / Q000235:genetics*; D015870:Gene Expression; D015972:Gene Expression Regulation, Neoplastic; D015723:Gene Library; D016147:Genes, Tumor Suppressor; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D017430:Prostate-Specific Antigen / Q000235:genetics; D011506:Proteins / Q000235:genetics; D012334:RNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification; D011990:Receptors, Transferrin / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D009418:S100 Proteins / Q000235:genetics; D015843:Serpins / Q000235:genetics; D014407:Tumor Cells, Cultured
D018076:DNA, Complementary; D011506:Proteins; D012334:RNA, Neoplasm; D011990:Receptors, Transferrin; D009418:S100 Proteins; C085344:SERPIN-B5; D015843:Serpins; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D017430:Prostate-Specific Antigen