Methods to isolate gene coding and flanking dna

C - Chemistry – Metallurgy – 12 – N

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C12N 15/10 (2006.01) C12N 15/63 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2375276

The invention provides methods for obtaining gene coding fragments, flanking fragments, or both gene coding and flanking fragments that can be readily characterized and used for a variety of purposes. The method involves preparing a genomic library comprising genomic DNA fragments, and hybridizing one or more nucleic acid primers, selected from a full-length cDNA, a 5' cDNA end, a 3'cDNA end, or a combination thereof, or a full- length mRNA, or portion thereof, or an RNA fragment, or a combination thereof, to the population of single stranded DNAs. A second strand is synthesized using a nucleic acid polymerase and the hybridized one or more nucleic acid primers, and a double stranded nucleic acid is produced. Any single stranded nucleic acid is removed and the nucleic acid reconstituted to produce a vector. This method is suitable for high throughput preparation and analysis of gene coding regions or flanking regions that can be used for preparing DNA arrays. Therefore, the present invention also provides arrays comprising flanking fragments, or flanking fragments attached to coding fragments. The present invention also pertains to promoter sequence tags, and 3' sequence tags.

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