Nucleic acid amplification in the presence of modified randomers

Abstract

The present invention is directed to a composition comprising a DNA Polymerase which is preferably thermostable, Deoxynucleotides, at least one primer oligonucleotide or a pair of amplification primers, and randomized 5-8 mer oligonucleotide, characterized in that said oligonucleotide comprises a modification with an organic hydrophobic moiety Such a composition is specifically useful for performing hot start PCR.

1 . A composition comprising
 a DNA polymerase,   deoxynucleotides,   at least one primer oligonucleotide, and   a randomized 5-8 mer oligonucleotide, wherein said oligonucleotide comprises a modification with an organic hydrophobic moiety.   
     
     
         2 . The composition according to  claim 1  wherein said modification is positioned at the 5′ end of said randomized oligonucleotide. 
     
     
         3 . The composition according to  claim 1  wherein said moiety is a pyrene or a stilbene. 
     
     
         4 . The composition according to  claim 1  wherein
 the DNA polymerase is thermostable and   the composition further comprises a pair of amplification primers.   
     
     
         5 . The composition according to  claim 1  further comprising a target nucleic acid sample. 
     
     
         6 . A kit comprising
 a DNA polymerase and   a randomized 5-8 mer oligonucleotide, wherein said oligonucleotide comprises a modification with an organic hydrophobic moiety.   
     
     
         7 . The kit according to  claim 6 , wherein said DNA polymerase is thermostable, the kit further comprising a pair of amplification primers. 
     
     
         8 . A method for amplification of a specific target nucleic acid comprising the steps of
 providing a sample suspected of containing the target nucleic acid,   adding a composition according to  claim 1 , and   performing at least a first primer extension reaction.   
     
     
         9 . A method for amplification of a specific target nucleic acid comprising the steps of
 providing a sample suspected of containing the target nucleic acid,   adding a composition according to  claim 4 ,   performing at least a first primer extension reaction, and   performing a nucleic acid amplification reaction.   
     
     
         10 . The method according to  claim 9 , wherein said nucleic acid amplification reaction is a polymerase chain reaction which is monitored in real time. 
     
     
         11 . The method according to  claim 8  wherein an amplification product generated by said amplification is subjected to a melting curve analysis.