Improvements in Bacterial Primers to Enhance Selective SSU rRNA Gene Amplification of Plant-associated Bacteria by Applying the LNA Oligonucleotide-PCR Clamping Technique

Microbes and Environments
Makoto IkenagaMasao Sakai

Abstract

PCR clamping by locked nucleic acid (LNA) oligonucleotides is an effective technique for selectively amplifying the community SSU rRNA genes of plant-associated bacteria. However, the original primer set often shows low amplification efficiency. In order to improve this efficiency, new primers were designed at positions to compete with LNA oligonucleotides. Three new sets displayed higher amplification efficiencies than the original; however, efficiency varied among the primer sets. Two new sets appeared to be available in consideration of bacterial profiles by next-generation sequencing. One new set, KU63f and KU1494r, may be applicable to the selective gene amplification of plant-associated bacteria.

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Citations

Jun 27, 2019·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Alfonso Soler-BistuéMarcelo E Tolmasky
Nov 14, 2019·Journal of Bioscience and Bioengineering·Xuan YinKazunori Takamine
Jul 27, 2021·Science China. Life Sciences·Liying ChenLili Zhang
Jan 13, 2022·The Science of the Total Environment·Sen LiGuanghua Wang

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Datasets Mentioned

BETA
PRJDB6970

Methods Mentioned

BETA
PCR

Software Mentioned

Qiime
ROSE

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