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Characterization of Non-Crosshybridizing DNA Oligonucleotides Manufactured In Vitro

Characterization of Non-Crosshybridizing DNA Oligonucleotides Manufactured In Vitro. J. Chen, R. Deaton, M. Garzon, J. W. Kim, D. Wood, H. Bi, D. Carpenter, Y.-Z. Wang DNA10, pp. 132-141 2004. 6. 29. MEC Seminar Summarized by In-Hee Lee. Introduction.

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Characterization of Non-Crosshybridizing DNA Oligonucleotides Manufactured In Vitro

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  1. Characterization of Non-Crosshybridizing DNA Oligonucleotides Manufactured In Vitro J. Chen, R. Deaton, M. Garzon, J. W. Kim, D. Wood, H. Bi, D. Carpenter, Y.-Z. Wang DNA10, pp. 132-141 2004. 6. 29. MEC Seminar Summarized by In-Hee Lee

  2. Introduction • In vitro protocol for generating non-crosshybridizing DNA oligonucleotides. • Refer to DNA8 proceedings.

  3. Introduction • Advantages • The oligonucleotides are selected in the conditions under which computations will be done. • The potential for very large libraries. • Prove the non-crosshybridizing characteristics of the library experimentally.

  4. Cloning and Sequencing of Library Oligonucleotides Decrease in the # of stable crosshybridization(?)

  5. Library Characterization with Gels • Its intensity means the number of extended products. • The intensity increase with cycle. • More product is amplified over the cycle.

  6. Library Characterization with Gels • No self-hybridization in top and bottom strands.

  7. Library Characterization with Spectroscopy • Melting curve investigation • Protocol product • Random DNA samples of length 20. • As a starting population of the protocol. • 40 non-crosshybridizing oligonucleotides of length 20 [Deaton at al., 2003] • Single-stranded oligonucleotide of length 20. • 3 & 4 as a standard for non-crosshybridization. • Watson-Crick pair of 2 sequences from 3. • As a standard for hybridization.

  8. Random seq. Top strands from cycle 4. NCH sequences Top and bottom strands from cycle 4. WC complements

  9. Conclusion • The experimental result indicate that the protocol was selecting for populations of non-crosshybridizing sequences.

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