Judith Ann St John

US Patent:

20080254516, Oct 16, 2008

Filed:

Mar 1, 2005

Appl. No.:

10/591051

Inventors:

Judith St. John - Littleton CO, US
Thomas W. Quinn - Westminster CO, US

International Classification:

C12P 19/34
C07H 21/00

US Classification:

435 912, 536 254, 435 911, 536 231, 536 2533

Abstract:

The invention provides efficient methods of isolating specific nucleic acid targets to obtain information from target nucleic acid sequences in a relatively short time period. DNA or cDNA is enzymatically digested into smaller fragments, double-stranded DNA linkers are added onto the ends of the DNA fragments to flank each fragment with a known DNA sequence. The fragments are mixed with an oligonucleotide probe that is bound to a marker and contains a conserved nucleic acid sequence of interest. The fragments that hybridize to the probe through nucleotide base pair complementation become indirectly connected to the marker. These target fragments are captured using a capture agent that specifically recognizes the marker and treated to prevent non-specific binding. Captured fragments are typically cloned prior to sequencing. The captured fragments may also be amplified using PCR to increase the efficiency of the cloning.