Parallel DNA polymerase chain reaction: Synthesis of two different PCR products from a DNA template

Abstract

Conventionally, in a polymerase chain reaction (PCR), oligonucleotide primers bind to the template DNA in an antiparallel complementary way and the template DNA is amplified as it is. Here we describe an approach in which the first primer binds in a parallel complementary orientation to the single-stranded DNA, leading to synthesis in a parallel direction. Further reactions happened in a conventional way leading to the synthesis of PCR product having polarity opposite to the template used. This is the first study showing that synthesis of DNA can happen also in a parallel direction. We report that from a single-stranded DNA template, two different but related PCR products can be synthesized.

Villain of Molecular Biology: Why are we not reproducible in research?

“…There are few reports in favour of parallel stranded DNA which summarize that there is no drastic difference in parallel and antiparallel DNA having mixed AT/GC composition 42 – 47 . Recently we have developed a PD-PCR technology based on parallel stranded DNA and we have concluded that two PCR products can be synthesized from a single stranded template DNA, one by conventional PCR and another by our approach 48 . In 2008, Lestienne et al reported a novel property of TFO (Triplex forming oligonucleotides-known for transcription inhibition) that Triple helix primer (THP) bounded to the duplex DNA in a parallel orientation can initiate DNA synthesis by various DNA polymerases of phage, retrovirus, bacteria and humans 49 , 50 .…”

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“…We strongly believe that errors reported in various bio-techniques can be resolved by considering both parallel and anti-parallel hybridization of DNA (24). Recently, we have developed a PD-PCR technique by applying the knowledge of both parallel and anti-parallel hybridization of DNA (25). In 2008, Lestienne et al has also reported that Triple helix primer (THP) bounded to the duplex DNA in a parallel orientation can initiate DNA synthesis (26).…”