公开(公告)号：US20220002797A1

公开(公告)日：2022-01-06

申请号：US17281269

申请日：2019-08-13

申请人： MAX-DELBRÜCK-CENTRUM FÜR MOLEKULARE MEDIZIN IN DER HELMHOLTZ-GEMEINSCHAFT

发明人： IVANO LEGNINI ,  JONATHAN ALLES ,  NIKOLAOS KARAISKOS ,  SALAH AYOUB ,  NIKOLAUS RAJEWSKY

IPC分类号： C12Q1/6869 ,  C12Q1/6806

摘要： The present disclosure provides a method for analyzing a polyribonucleotide, wherein the polyribonucleotide is an mRNA molecule with a poly(A) tail, said method comprising: (a) obtaining a labeled polyribonucleotide by GI tailing; followed by (b) providing a second molecule comprising a first primer recognition sequence followed by a sequence of C nucleotide residues linked to a sequence of T nucleotide residues; followed by (c) obtaining a complex of said labeled polyribonucleotide and said second molecule; followed by (d) obtaining an extended second molecule by (d1) extending the 3′ end of the second molecule by synthesizing a sequence that is complementary to the labeled polyribonucleotide; followed by (d2) extending the 3′ end of the second molecule by adding at least 1 to 5 C nucleotide residues, followed by a second primer recognition sequence; said method providing information on entire polyribonucleotides, in particular mRNA isoforms and their respective poly(A) tails.