Dual asymmetric PCR: one-step construction of synthetic genes.
GS Sandhu, RA Aleff, BC Kline - Biotechniques, 1992 - europepmc.org
GS Sandhu, RA Aleff, BC Kline
Biotechniques, 1992•europepmc.orgWe have developed a one-step process for constructing synthetic genes. Four adjacent
oligonucleotides 17-100 bases in length having short overlaps of 15-17 bases are used as
primers in a PCR mixture. The quantity of the two internal primers is highly limited, and the
resultant reaction causes an asymmetric single-stranded amplification of the two halves of
the total sequence due to an excess of the two flanking primers. In subsequent PCR cycles,
these dual asymmetrically amplified fragments, which overlap each other, yield a double …
oligonucleotides 17-100 bases in length having short overlaps of 15-17 bases are used as
primers in a PCR mixture. The quantity of the two internal primers is highly limited, and the
resultant reaction causes an asymmetric single-stranded amplification of the two halves of
the total sequence due to an excess of the two flanking primers. In subsequent PCR cycles,
these dual asymmetrically amplified fragments, which overlap each other, yield a double …
We have developed a one-step process for constructing synthetic genes. Four adjacent oligonucleotides 17-100 bases in length having short overlaps of 15-17 bases are used as primers in a PCR mixture. The quantity of the two internal primers is highly limited, and the resultant reaction causes an asymmetric single-stranded amplification of the two halves of the total sequence due to an excess of the two flanking primers. In subsequent PCR cycles, these dual asymmetrically amplified fragments, which overlap each other, yield a double-stranded, full-length product.
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