US2009280498A1PendingUtilityA1

Rapid and efficient capture of dna from sample without using cell lysing reagent

Individually held — no corporate assignee on recordPriority: May 4, 1999Filed: Jul 15, 2009Published: Nov 12, 2009

Est. expiryMay 4, 2019(expired)· nominal 20-yr term from priority

Inventors:Robert T. Belly Jianbo Sun

C12Q 2600/156C12Q 1/6806C12N 15/1003C12Q 1/6886C12Q 2600/16

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Cited by

References

Claims

Abstract

Nucleic acids can be made available for amplification or other treatment after admixture of a sample with specific weakly basic polymers to form a precipitate with the nucleic acids at acidic pH. After removing non-precipitated materials, the pH is then made basic, thereby releasing the nucleic acids from the polymer. This method for preparing specimen samples is simple and quite rapid, and the released nucleic acids can be further treated in hybridization assays or amplification procedures. No surfactant or other cell lysing reagents are employed. The weakly basic polymers are water-soluble and cationic at acidic pH, but neutral in charge at basic pH.

Claims

exact text as granted — not AI-modified

1 . A kit for detection of K-ras mutation in a biological sample, said kit comprising a diagnostic K-ras primer selected from the group consisting of: 
       
         
           
                 
                 
               
                   &lt;SEQ ID: NO 1&gt; 
                     
                 
                 
                 
                 
               
                     
                   (a) TGAATATAAA CTTGTGGTAC CTGGAGC T (5K15S), 
                     
                 
                     
                     
                 
                 
                 
               
                   &lt;SEQ ID: NO 2&gt; 
                     
                 
                 
                 
                 
               
                     
                   (b) ATATAAACTT GTGGTAGTTC CAGCTGGT (5K37), 
                     
                 
                     
                     
                 
                 
                 
               
                   &lt;SEQ ID: NO 3&gt; 
                     
                 
                 
                 
                 
               
                     
                   (c) GAATTAGCTG TATCGTCAAG GCACTC (3K42), 
                     
                 
                     
                     
                 
                 
                 
               
                   &lt;SEQ ID: NO 4&gt; 
                     
                 
                 
                 
                 
               
                     
                   (d) TCAGCAAAGA CAAGACAGGT A (5BK5), 
                     
                 
                     
                     
                 
                 
                 
               
                   &lt;SEQ ID: NO 5&gt; 
                     
                 
                 
                 
                 
               
                     
                   (e) TATAGATGGT GAAACCTGTT TGTTGG (5N12A), 
                     
                 
                     
                     
                 
                 
                 
               
                   &lt;SEQ ID: NO 6&gt; 
                     
                 
                 
                 
                 
               
                     
                   (f) CTTGCTATTA TTGATGGCAA CCACACAGA (3N13A) 
                     
                 
             
                
               
            
             
                
                
               
            
             
                
               
            
             
                
                
               
            
             
                
               
            
             
                
                
               
            
             
                
               
            
             
                
                
               
            
             
                
               
            
             
                
                
               
            
             
                
               
            
             
                
               
            
           
         
         (g) any combination of the foregoing. 
       
     
     
         2 . The oligonucleotide TGAATATAAA CTTGTGGTAC CTGGAGC T &lt;SEQ ID: NO 1&gt;. 
     
     
         3 . The oligonucleotide ATATAAACTT GTGGTAGTTC CAGCTGGT &lt;SEQ ID: NO 2&gt;. 
     
     
         4 . The oligonucleotide GAATTAGCTG TATCGTCAAG GCACTC &lt;SEQ ID: NO 3&gt;. 
     
     
         5 . The oligonucleotide TCAGCAAAGA CAAGACAGGT A &lt;SEQ ID: NO 4&gt;. 
     
     
         6 . The oligonucleotide TATAGATGGT GAAACCTGTT TGTTGG &lt;SEQ ID: NO 5&gt;. 
     
     
         7 . The oligonucleotide CTTGCTATTA TTGATGGCAA CCACACAGA &lt;SEQ ID: NO 6&gt;. 
     
     
         8 . A K-ras diagnostic primer comprising the oligonucleotide TGAATATAAA CTTGTGGTAC CTGGAGC T&lt;SEQ ID: NO 1&gt;. 
     
     
         9 . A K-ras diagnostic primer comprising the oligonucleotide ATATAAACTT GTGGTAGTTC CAGCTGGT &lt;SEQ ID: NO 2&gt;. 
     
     
         10 . A K-ras diagnostic primer comprising the oligonucleotide GAATTAGCTG TATCGTCAAG GCACTC &lt;SEQ ID: NO 3&gt;. 
     
     
         11 . A K-ras diagnostic primer comprising the oligonucleotide TCAGCAAAGA CAAGACAGGT A &lt;SEQ ID: NO 4&gt;. 
     
     
         12 . A K-ras diagnostic primer comprising the oligonucleotide TATAGATGGT GAAACCTGTT TGTTGG &lt;SEQ ID: NO 5&gt;. 
     
     
         13 . A K-ras diagnostic primer comprising the oligonucleotide CTTGCTATTA TTGATGGCAA CCACACAGA &lt;SEQ ID: NO 6&gt;.

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