| 摘要 |
A method for amplification and capture of nucleic acid sequences comprising, annealing a forward primer to a nucleic acid in a first reaction vessel comprising fewer than four different dNTPs; extending the forward primer with the dNTPs to form an terminated-extended primer; releasing the extended primer; exponentially amplifying the terminated-extended primer in a second reaction vessel that includes a reverse primer, four different dNTPs and a capture probe comprising n oligonucleotides, wherein fewer than n of the oligonucleotides are locking nucleic acids; and concurrently capturing one of the extended primers in the second reaction vessel with the capture probe while amplifying the extended primer. In alternative embodiments, annealing, extending and releasing occur at a first reaction temperature that is substantially isothermal and in the absence of a helicase. In addition, the steps of exponentially amplifying and capturing occur at a second reaction temperature that is substantially isothermal.
|
