Subtractive hybridization of cDNA from small amounts of plant tissue.
A subtractive hybridization method is described that allows the generation of a subtractive gene library from small amounts of plant or other eukaryotic tissues. The method uses paramagnetic oligo-dT beads to capture poly-adenylated mRNA and to synthesize the complementary cDNA on a solid support. The use of magnetic beads facilitates the change of reaction buffers and the removal of primers and minimizes yield losses. Subtracted material obtained from this method can either be cloned directly or used to screen a specific library.
Heinrich
T
T
Western Australian State Agricultural Biotechnology Centre, Murdoch University, Perth, Australia. tatjana@central.murdoch.edu.au
Washer
S
S
Marshall
J
J
Jones
M G
MG
Potter
R H
RH
D018076:DNA, Complementary / Q000235:genetics*; D018744:DNA, Plant / Q000235:genetics*; D018551:Solanum lycopersicum / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D011198:Solanum tuberosum / Q000235:genetics*; D013382:Subtraction Technique
D018076:DNA, Complementary; D018744:DNA, Plant