Details
| Original language | English |
|---|---|
| Pages (from-to) | 953-958 |
| Number of pages | 6 |
| Journal | Engineering in life sciences |
| Volume | 17 |
| Issue number | 8 |
| Publication status | Published - 19 Jul 2017 |
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.
Keywords
- Aptamer, By-product formation, Emulsion PCR, Polymerase chain reaction, SELEX
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biotechnology
- Environmental Science(all)
- Environmental Engineering
- Chemical Engineering(all)
- Bioengineering
Cite this
- Standard
- Harvard
- Apa
- Vancouver
- BibTeX
- RIS
In: Engineering in life sciences, Vol. 17, No. 8, 19.07.2017, p. 953-958.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Comparing two conventional methods of emulsion PCR and optimizing of Tegosoft-based emulsion PCR
AU - Witt, Martin
AU - Phung, Ngoc Linh
AU - Stalke, Amelie
AU - Walter, Johanna Gabriela
AU - Stahl, Frank
AU - von Neuhoff, Nils
AU - Scheper, Thomas
N1 - Funding information: This project was funded by Biofabrication for NIFE (supported by the german Lower Saxonian Ministry for Science and Culture and the VolkswagenStiftung). Tegosoft DEC and ABIL WE 09 were kindly provided by Evonik Industries AG.
PY - 2017/7/19
Y1 - 2017/7/19
N2 - 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.
AB - 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.
KW - Aptamer
KW - By-product formation
KW - Emulsion PCR
KW - Polymerase chain reaction
KW - SELEX
UR - http://www.scopus.com/inward/record.url?scp=85028352121&partnerID=8YFLogxK
U2 - 10.1002/elsc.201700047
DO - 10.1002/elsc.201700047
M3 - Article
C2 - 32624844
AN - SCOPUS:85028352121
VL - 17
SP - 953
EP - 958
JO - Engineering in life sciences
JF - Engineering in life sciences
SN - 1618-0240
IS - 8
ER -