Removal of known, abundant cDNA species by specific double-stranded cDNA synthesis-based subtraction.
An efficient and simple method for removing known, abundant cDNA species from the cDNA pool of highly differentiated cells is reported. The method involves preparation of sscDNA, followed by dscDNA-synthesis of known, abundant cDNA species led by specific primers and removal of the synthesized dscDNA with hydroxyapatite. By using this method, the globin cDNAs were reduced to less than 10(-5) of their original abundance. The results suggest that this method may facilitate the isolation of new genes from specific cells or tissues.
Yang
X O
XO
National Laboratory of Medical Molecular Biology, Peking Union Medical College, Beijing, P.R. China.
Huang
X D
XD
Zhao
H L
HL
Zhang
J W
JW
D001483:Base Sequence; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods; D013382:Subtraction Technique
D017931:DNA Primers; D018076:DNA, Complementary