Parallel DNA polymerase chain reaction: Synthesis of …
If a person were to attempt extending a synthetic oligonucleotide prepared to be complementary to a target on human DNA by just one base, using DNA polymerase and dideoxynucleoside triphosphates, using four different tubes each containing all four bases, but only one of them in each tube alpha-labeled with 32P, optimistically one might be able to discover the identity of the nucleotide on the DNA target just three-prime of the oligomer. Dideoxy-sequencing worked that way…but…Huge but…that only worked on cloned DNA where the ratio of target to non-target DNA was increased by a factor of about a million. Fortunately for me I was thinking about other things that might go wrong than just the brute improbability that only the right sequence would be engaged. I paid just enough attention to this hypothetical problem to plan on using two oligonucleotides, one designed for each strand of the target sequence coming at the base pair in question from either side. Although these two sides would be far distant in the denatured reaction mixture they would still represent complementary strands and if one told me that a 'T' was three-prime to one oligo, the other should have told me 'A' was three-prime to the other. Not much of a control, but I had oligos to burn. In fact that was what I was trying to do. We had excess oligos on our hands.
of polymerase chain reactions using ..
Polymerase chain reaction - Wikipedia
Dr. Mullis received a Nobel Prize in chemistry in 1993, for his invention of the polymerase chain reaction (PCR). The process, which Dr. Mullis conceptualized in 1983, is hailed as one of the monumental scientific techniques of the twentieth century. A method of amplifying DNA, PCR multiplies a single, microscopic strand of the genetic material billions of times within hours. The process has multiple applications in medicine, genetics, biotechnology, and forensics. PCR, because of its ability to extract DNA from fossils, is in reality the basis of a new scientific discipline, paleobiology.
Reverse transcription polymerase chain reaction - Wikipedia
DNA strands containing an unnatural T-triazole-T linkage have been synthesized by click DNA ligation between oligonucleotides with 3′-AZT and 5′-propargylamido dT and amplified efficiently by polymerase chain reaction (PCR) using several different polymerases. DNA sequencing of PCR amplicons and clones in two different sequence contexts revealed the presence of a single thymidine at the ligation site. The remarkable ability of thermostable polymerases to reproducibly copy DNA templates containing such an unnatural backbone opens up intriguing possibilities in gene synthesis, genetic analysis, biology, and nanotechnology.