Method for the 5'-cap-dependent amplification of cDNAs
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May 1, 2003
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May 5, 1999
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Abstract
The invention concerns a method for the modification, cloning and amplification of cDNAs which are complete at their 5' end which is essentially characterized in that the first strand cDNA synthesis is carried out in the presence of manganese.sup.2+ ions or manganese.sup.2+ is added as an additive at a later time. The CAP structure at the 5' end of the reversely transcribed mRNA triggers the attachment of deoxy-cytosines to the 3' end of the cDNA with high efficiency. In a preferred embodiment a controlled ribonucleotide tailing is carried out with the aid of terminal transferase following the first strand cDNA synthesis.
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