1. Summarized by In-Hee Lee
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
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
Cycle # %
Start
2(2)
20(20) 1
8(3)
22(8.3) 2
12(4)
33(11) 3
5(3)
23.8(14.3) 4
6(3)
28.6(14.3)
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.