DNA-SEQUENCING BY HYBRIDIZATION TO OLIGONUCLEOTIDE MATRIX - CALCULATION OF CONTINUOUS STACKING HYBRIDIZATION EFFICIENCY

被引:22
作者
LYSOV, YP [1 ]
CHERNYI, AA [1 ]
BALAEFF, AA [1 ]
BEATTIE, KL [1 ]
MIRZABEKOV, AD [1 ]
机构
[1] HOUSTON ADV RES CTR,DNA TECHNOL LAB,THE WOODLANDS,TX 77380
关键词
D O I
10.1080/07391102.1994.10508033
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
In this paper we consider the efficiency of additional rounds of ''continuous stacking'' hybridization in DNA sequence reconstruction by hybridization with oligonucleotide matrix (SHOM). After the initial hybridization of target DNA with the matrix of oligonucleotides of fixed length L some additional hybridizations should be carried out in the presence of fluorescently labeled oligonucleotides of another length l. These additional oligonucleotides can hybridize in tandem with matrix tuples (continuous stacking hybridization) thus forming an extended duplex with the target DNA strand. The additional data obtained allows resolutions of branching points arising in the reconstruction procedure. Multiple rounds of continuous stacking hybridization considerably increase the efficiency of the sequencing method, eventually approaching the power of (L+l)-matrix. We develop here an; algorithm that allows us to minimize the number of additional hybridization steps, by assembling sets of I-tuples to be added together in each round of continuous stacking hybridization. For SHOM using a matrix of octanucleotides, continuous stacking hybridization with pentanucleotides increases the length of unambiguously sequenced DNA from 200 to several thousands of base pairs.
引用
收藏
页码:797 / 812
页数:16
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