A RAPID ISOLATION OF THE UNKNOWN 5'-FLANKING SEQUENCE OF HUMAN CENP-B CDNA WITH POLYMERASE CHAIN-REACTIONS

作者：SUGIMOTO K^*; HIMENO M

来源：Agricultural and Biological Chemistry, 1991, 55(11): 2687-2692.

DOI：10.1080/00021369.1991.10871042

We rapidly and efficiently isolated the 5'-region of cDNA encoding the N-terminal region of human centromere antigen B (CENP-B) including an ATG methionine codon by polymerase chain reactions (PCR). The unknown 5'-flanking sequence of the cDNA was amplified using an adaptor-sequence ligated to the 5' end as a universal primer sequence. To locate the target fragments, we did an additional PCR with another set of two internal primers using samples of the size-fractionated products as templates, rather than using the conventional hybridization procedure. This approach can further be applied to the analysis of other unknown flanking sequences of cDNA or genomic DNA.

• 出版日期1991-11