Shot-gun sequencing and amplification without cloning

Details Shot-gun sequencing and amplification without cloning Shot-gun sequencing and amplification without cloning

2006-06-06

2006-06-06

Lu, Frank W. (Department: 1634)

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Preparing compound containing saccharide radical

C435S006120, C435S091100, C436S094000, C536S023100, C536S024300, C536S024330

Reexamination Certificate

active

07056706

ABSTRACT:
Disclosed is a method for sequencing and amplifying nucleic acid templates wherein a degenerate primer with a fixed sequence region and a random sequence region is utilized. By determining the statistical expectancy of the fixed sequence in the nucleic acid template, this determines the average length of a nucleic acid template that can be sequenced. During the annealing of such a primer with the nucleic acid template, the fixed sequence determines where the complete primer binds by binding to its complementary sequence on the nucleic acid template. The random sequence regions of the primers make it possible for the presence of a unique sequence adjacent to the fixed sequence to be present, thus providing a primer with full complementarity with the nucleic acid template. Thus, this procedure is able to provide a full-length primer with a fully complementary sequence capable of binding statistically once within an expected length of the nucleic acid template, even though the sequence of the template is unknown. The method can also be adopted for use in PCR amplification of a nucleic acid template.

REFERENCES:
patent: 5807679 (1998-09-01), Kamb
patent: 6090552 (2000-07-01), Nazarenko et al.
patent: 6521428 (2003-02-01), Senapathy
Sommer et al., Minimal homology requirement for PCR primers. Nucleic Acids Research, 17, 6749, 1989.

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