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Amplified fluorescence detection of DNA and RNA hybridization by surface initiated enzymatic polymerization (SIEP)

Publication ,  Journal Article
Tjong, V; Yu, H; Hucknall, A; Chilkoti, A
Published in: Technical Proceedings of the 2013 NSTI Nanotechnology Conference and Expo, NSTI-Nanotech 2013
August 9, 2013

Detection of nucleic acid on-chip has been of great interest due to its potential as research tools and for diagnostic applications. We present a new on-chip, isothermal signal amplification technique for the direct detection of unmodified DNA and RNA on microarrays using terminal deoxy nucleotidyl transferase (TdT), a template-independent DNA polymerase that catalyzes the sequential addition of deoxynucleotides (dNTPs) at the 3'-OH group of a DNA primer and yeast poly(A) polymerase (PaP), an RNA polymerase that catalyzes polyadenylation at the 3'-OH an RNA primer. We utilized their ability to catalyze the formation of long polynucleotide and TdT's ability to incorporate unnatural nucleotide substrates such as fluorescent nucleotides (F-dNTP) into a long polymer chain of single stranded DNA (ssDNA) from the 3'-OH of a target DNA or RNA that is tethered on the surface through hybridization. We obtained dose reponse curves of hybridized DNA and RNA with LOD in -1-10 pM.

Duke Scholars

Published In

Technical Proceedings of the 2013 NSTI Nanotechnology Conference and Expo, NSTI-Nanotech 2013

Publication Date

August 9, 2013

Volume

3

Start / End Page

32 / 36
 

Citation

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ICMJE
MLA
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Tjong, V., Yu, H., Hucknall, A., & Chilkoti, A. (2013). Amplified fluorescence detection of DNA and RNA hybridization by surface initiated enzymatic polymerization (SIEP). Technical Proceedings of the 2013 NSTI Nanotechnology Conference and Expo, NSTI-Nanotech 2013, 3, 32–36.
Tjong, V., H. Yu, A. Hucknall, and A. Chilkoti. “Amplified fluorescence detection of DNA and RNA hybridization by surface initiated enzymatic polymerization (SIEP).” Technical Proceedings of the 2013 NSTI Nanotechnology Conference and Expo, NSTI-Nanotech 2013 3 (August 9, 2013): 32–36.
Tjong V, Yu H, Hucknall A, Chilkoti A. Amplified fluorescence detection of DNA and RNA hybridization by surface initiated enzymatic polymerization (SIEP). Technical Proceedings of the 2013 NSTI Nanotechnology Conference and Expo, NSTI-Nanotech 2013. 2013 Aug 9;3:32–6.
Tjong, V., et al. “Amplified fluorescence detection of DNA and RNA hybridization by surface initiated enzymatic polymerization (SIEP).” Technical Proceedings of the 2013 NSTI Nanotechnology Conference and Expo, NSTI-Nanotech 2013, vol. 3, Aug. 2013, pp. 32–36.
Tjong V, Yu H, Hucknall A, Chilkoti A. Amplified fluorescence detection of DNA and RNA hybridization by surface initiated enzymatic polymerization (SIEP). Technical Proceedings of the 2013 NSTI Nanotechnology Conference and Expo, NSTI-Nanotech 2013. 2013 Aug 9;3:32–36.

Published In

Technical Proceedings of the 2013 NSTI Nanotechnology Conference and Expo, NSTI-Nanotech 2013

Publication Date

August 9, 2013

Volume

3

Start / End Page

32 / 36