Patent ID: 6316229
Filing Date: 2001-11-13
Classification: C12Q

Abstract:
A method of amplifying nucleic acid sequences, the method comprising,(a) mixing a half probe with a target sample comprising a target sequence, to produce a probe-target mixture, and incubating the probe-target mixture under conditions that promote hybridization between the half probe and the target sequence in the probe-target mixture,(b) mixing a probe/primer with the probe-target mixture, and incubating the probe-target mixture under conditions that promote hybridization between the probe/primer and the target sequence,(c) mixing ligase with the probe-target mixture, to produce a ligation mixture, and incubating the ligation mixture under conditions that promote ligation of the half probe and the probe/primer to form a rolling circle replication primer,(d) mixing an amplification target circle with the rolling circle replication primer, to produce a primer-ATC mixture, and incubating the primer-ATC mixture under conditions that promote hybridization between the amplification target circle and the rolling circle replication primer in the primer-ATC mixture, and(e) mixing DNA polymerase with the primer-ATC mixture, to produce a polymerase-ATC mixture, and incubating the polymerase-ATC mixture under conditions that promote replication of the amplification target circle,wherein replication of the amplification target circle results in the formation of tandem sequence DNA.