Compounds for use in pcr systems and applications thereof

Abstract

This disclosure relates to novel compounds for use in various compositions, kits and methods, including, for example, use in polymerase storage buffers and in nucleic acid synthesis or amplification reactions such as a polymerase chain reaction (PCR). Methods for preparing the novel compounds are also described.

What is claimed is: 
     
         1 . A compound of Formula I: 
       
         
           
           
               
               
           
         
         wherein: 
         R 1  is H, (C 1 -C 30 ) alkyl, (C 1 -C 30 ) substituted alkyl, (C 1 -C 30 ) heteroalkyl, (C 1 -C 30 ) substituted heteroalkyl, aryl, heteroaryl, substituted heteroaryl, or phenyl; 
         R 2  is H, CH 3 , CH(CH 3 ) 2 , CH 2 (C 6 H 5 ), or C(CH 3 ) 3 ; 
         R 4  is H, CH 3 , CH(CH 3 ) 2 , C 6 H 5 , CH 2 (C 6 H 5 ), C(CH 3 ) 3 , CH 2 CH(CH 3 ) 2 , CHCH 2 CH(CH 3 ) 2 , CH 2 C 6 H 50 H, CH 2 C═CH NH(C 6 H 5 ), CH 2 C═CHN═CHNH, CH 2 COOH, CH 2 CONH 2 , (CH 2 ) 2 CONH 2 , (CH 2 ) 2 COOH, CH 2 SH, (CH 2 ) n NH, (CH 2 ) n N, CH 2 OH, CH(OH)CH 3 , (CH 2 ) 2 SCH 3 , (CH 2 ) 3 NHC(NH 2 )═NH; 
         R 3  together with R 5  forms a 5- or 6-membered ring which is optionally substituted with at least one (C 1 -C 30 ) alkyl, (C 1 -C 30 ) substituted alkyl, (C 1 -C 30 ) heteroalkyl, or (C 1 -C 30 ) substituted heteroalkyl; and 
         each n is from 1-30. 
       
     
     
         2 . The compound of  claim 1 , wherein R 1  is a C 8  alkyl, C 12  alkyl, or C 16  alkyl. 
     
     
         3 . The compound of  claim 1 , wherein R 2  is H or CH 3 . 
     
     
         4 . The compound of  claim 1 , wherein R 4  is H, (CH 2 ) n NH, or (CH 2 ) n N. 
     
     
         5 . The compound of  claim 1 , wherein R 3  together with R 5  forms a 5-membered ring. 
     
     
         6 . The compound of  claim 1 , wherein the compound of Formula I is: 
       
         
           
           
               
               
           
         
       
     
     
         7 . A composition comprising a polymerase and the compound of  claim 1 . 
     
     
         8 . The composition of  claim 7 , wherein said polymerase is a thermostable polymerase. 
     
     
         9 . The composition of  claim 7 , further comprising dNTPs, one or more primers, and one or more detectable labels. 
     
     
         10 . The composition of  claim 7 , further comprising a polymerase inhibitor. 
     
     
         11 . A method for increasing the efficiency of a polymerase, the method comprising the steps of:
 a) mixing a target nucleic acid with at least one polymerase, at least one primer, dNTPs, and the compound of  claim 1 ; and   b) amplifying said target nucleic acid.   
     
     
         12 . A method for detecting a target nucleic acid in a sample, the method comprising the steps of:
 a) forming a reaction mixture comprising at least one polymerase, at least one primer, dNTPs, the compound of  claim 1 , and a detectable label;   b) amplifying said target nucleic acid; and   c) detecting a signal generated from said detectable label indicative of the presence and/or amount of said target nucleic acid in said sample.   
     
     
         13 . A method for inhibiting inactivation of a polymerase in a thermal cycling process, said method comprising contacting said polymerase with the compound of  claim 1  during the thermal cycling process. 
     
     
         14 . A nucleic acid amplification reaction mixture comprising:
 a) at least one polymerase;   b) dNTPs; and   c) the compound according to  claim 1 .   
     
     
         15 . The nucleic acid amplification reaction mixture of  claim 14 , further comprising at least one primer. 
     
     
         16 . The nucleic acid amplification reaction mixture of  claim 14 , wherein said polymerase is thermostable. 
     
     
         17 . The nucleic acid amplification reaction mixture of  claim 14 , further comprising at least one antibody. 
     
     
         18 . The nucleic acid amplification reaction mixture of  claim 14 , further comprising at least one oligonucleotide polymerase inhibitor.