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METHOD OF AMPLIFYING MRNAS AND FOR PREPARING FULL LENGTH MRNA LIBRARIES
METHOD OF AMPLIFYING MRNAS AND FOR PREPARING FULL LENGTH MRNA LIBRARIES
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机译：扩增mRNA的方法和制备全长mRNA文库
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摘要
The inventive method for amplifying at least one RNA which is contained in a sample includes reverse transcribing the at least one RNA using a first primer, adding a dideoxy nucleotide which is modified in the 3'-position with a first partner of a pair of azide and alkyne molecules by action of a template-independent polymerase to attach a single 3'-azide- or 3'-alkyne-modified dideoxy nucleotide at the 3'-end of the obtained cDNA, adding an adapter molecule which comprises a polynucleotide sequence and at its 5'-end a second partner of such pair of azide and alkyne molecules and ligating the adapter to the 3'-modified cDNA under formation of a triazole linkage, adding a second primer which is complementary to at least a part of the adapter molecule and which contains at its 3'-end a nucleotide which is complementary to the dideoxy nucleotide at the 3'-end of the cDNA to effect hybridization and binding of the second primer overlapping the triazole linkage, adding a third primer and amplifying the full length cDNA.;Uses of such method especially for preparing a full length RNA library and for sequencing of a plurality of RNAs contained in a sample, as well as reagent kits for perfoming such methods are also disclosed and included in the invention..
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