An Alternative Hot Start PCR Method Using a Nuclease-Deficient ExoIII from Escherichia coli.

The Hot Start polymerase chain reaction (Hot Start PCR) is designed to reduce off-target amplification by blocking DNA polymerase extension at room temperature until the desired temperature is reached. In this study, we investigated a new method of Hot Start PCR that uses a modified Escherichia coli Exonuclease III (EcoExoIIIM) by substituting residues in the DNA-binding pocket and catalytic center. The results showed that PCR amplification yield and specificity were significantly promoted by the addition of EcoExoIIIM. We hypothesize that non-specific binding of primers at room temperature is prevented by binding of the primed template by EcoExoIIIM, which is then released from the DNA by heat denaturation before the first PCR cycle. Through this mechanism, PCR would be enhanced by reducing off-target extension at room temperature

An Alternative Hot Start PCR Method Using a Nuclease-Deficient ExoIII from Escherichia coli

Abstract(#br)The Hot Start polymerase chain reaction (Hot Start PCR) is designed to reduce off-target amplification by blocking DNA polymerase extension at room temperature until the desired temperature is reached. In this study, we investigated a new method of Hot Start PCR that uses a modified Escherichia coli Exonuclease III (EcoExoIIIM) by substituting residues in the DNA-binding pocket and catalytic center. The results showed that PCR amplification yield and specificity were significantly promoted by the addition of EcoExoIIIM. We hypothesize that non-specific binding of primers at room temperature is prevented by binding of the primed template by EcoExoIIIM, which is then released from the DNA by heat denaturation before the first PCR cycle. Through this mechanism, PCR would be..

The complete chloroplast genome of Siraitia grosvenorii

Siraitia grosvenorii belongs to Cucurbitaceae. It is widely used in beverage raw materials and traditional Chinese medicine. In this study, Illumina sequencing method was used to establish its complete chloroplast (cp) genome. The complete cp genome is 157,132 bp in length, with a large single copy region (LSC) of 92,442 bp and a small single copy region (SSC) of 21,232 bp, which were separated by a pair of inverted repeat (IR) regions of 21,729 bp. The complete cp genome consists of 85 coding sequences (CDS), 39 tRNA, and eight rRNA genes. Phylogenetic analysis showed that S. grosvenorii was clustered into Cucurbitaceae

Concise Formal Synthesis of (+)-Neopeltolide

A concise formal synthesis of (+)-neopeltolide (<b>1</b>) has been accomplished. The synthesis demonstrated high atom efficiency employing only one step of functional group protection. Key steps involved iridium-catalyzed double asymmetric carbonyl allylation, palladium-catalyzed intramolecular alkoxycarbonylation, ruthenium-catalyzed olefin isomerization, and ring-closing metathesis