Patent ID: 6368791
Filing Date: 2002-04-09
Classification: C12Q

Abstract:
A method of amplifying an unknown region of a translocation partner which flanks a section of the breakpoint cluster region of a leukemia-associated MLL gene, the method comprising:(a) providing a template polynucleotide comprising a sense strand which comprises the section and the unknown region, wherein the unknown region is nearer the 3â€² end of the sense strand than is the section, wherein the section comprises a first portion and a second portion, and wherein the first portion is nearer the unknown region than is the second portion; (b) ligating a loop-forming oligonucleotide to the 3â€²-end of the sense strand, wherein the loop-forming oligonucleotide is complementary to the first portion; (c) annealing the loop-forming oligonucleotide with the first portion to generate a panhandle structure; (d) subjecting the panhandle structure to extension, whereby an additional region complementary to the second portion is generated at the free end of the loop-forming oligonucleotide; and (e) subjecting the panhandle structure to PCR in the presence of a first primer homologous with the second portion, whereby the unknown region is amplified.