ASYMMETRICAL PCR AMPLIFICATION

摘要

A DNA amplification approach, enabling the amplification of DNA targets with unknown sequences, involves successive PCR steps. In a first reaction, linear amplification of the target nucleic acid is achieved using one or more specially designed primers. In the second reaction, additional primers are added and exponential amplification of a double stranded DNA is achieved. Combined with conventional DNA sequencing procedures, this approach can be used for the direct sequencing of genomic DNA, avoiding the need to sub-clone large fragments. With the inventive method, moreover, sequencing information can be gathered in a much less costly and labor-intensive manner than was possible previously.