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Comparing two conventional methods of emulsion PCR and optimizing of Tegosoft-based emulsion PCR.

ABSTRACT: The selection of aptamers represents a promising route in the development of high affinity ligands. In these processes the formation of by-products is a common problem during the PCR-based amplification of complex oligonucleotide libraries. One approach to overcome this drawback is to separate each template oligonucleotide into an individual reaction compartment provided by a droplet. This method, termed emulsion PCR (ePCR), has already emerged to a standard method in sample preparation for 2nd generation sequencing. In this work, we compare different literature protocols that have been developed to generate stable emulsions for ePCR. We investigate different emulsification methods and evaluate the importance of the initial template concentration. We demonstrate that emulsion stability is of utmost importance for the successful inhibition of by-product formation and give an optimized protocol for generation of an emulsified PCR.

SUBMITTER: Witt M

PROVIDER: S-EPMC6999282 | biostudies-literature | 2017 Aug

REPOSITORIES: biostudies-literature

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Comparing two conventional methods of emulsion PCR and optimizing of Tegosoft-based emulsion PCR.

Witt Martin M Phung Ngoc Linh NL Stalke Amelie A Walter Johanna-Gabriela JG Stahl Frank F von Neuhoff Nils N Scheper Thomas T

Engineering in life sciences 20170828 8

The selection of aptamers represents a promising route in the development of high affinity ligands. In these processes the formation of by-products is a common problem during the PCR-based amplification of complex oligonucleotide libraries. One approach to overcome this drawback is to separate each template oligonucleotide into an individual reaction compartment provided by a droplet. This method, termed emulsion PCR (ePCR), has already emerged to a standard method in sample preparation for 2nd ...[more]

PMID: 32624844

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Comparing two conventional methods of emulsion PCR and optimizing of Tegosoft-based emulsion PCR.

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Comparing two conventional methods of emulsion PCR and optimizing of Tegosoft-based emulsion PCR.

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