Construction of subtractive cDNA libraries from limited amounts of mRNA and multiple cycles of subtraction.
A new strategy to construct subtractive cDNA libraries was developed. The method was based on multiple subtraction cycles from small amounts of RNA using paramagnetic technology. An additional application of this technique is the potential to recover (and reuse) the same mRNAs or the single-stranded cDNA from the solid paramagnetic support. The result was assessed in the generation of a specific prepuberal mouse testis library.
López-Fernández
L A
LA
Centro de Investigaciones Biológicas, C.S.I.C., Madrid, Spain.
del Mazo
J
J
D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000302:isolation & purification / Q000378:metabolism; D015723:Gene Library; D008280:Magnetics; D008297:Male; D051379:Mice; D008863:Microspheres; D009691:Nucleic Acid Denaturation; D009693:Nucleic Acid Hybridization; D011061:Poly A / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D012313:RNA / Q000302:isolation & purification; D012333:RNA, Messenger / Q000302:isolation & purification / Q000378:metabolism; D013737:Testis / Q000737:chemistry
D018076:DNA, Complementary; D012333:RNA, Messenger; D011061:Poly A; D012313:RNA