Amplification of nucleic acids

Abstract

Provided herein is technology relating to amplification of nucleic acids and particularly, but not exclusively, to compositions and methods for doing improving the polymerase chain reaction and providing reagents for polymerase chain reaction with improved stability.

We claim: 
     
         1 . A method of detecting a nucleic acid in a realtime PCR composition, said method comprising providing a reaction composition comprising a polymerase, a first primer, a second primer, nucleotides, and 0.0001 to 0.01% (w/w) MIT. 
     
     
         2 . The method of  claim 1 , wherein said MIT is a PCR enhancer. 
     
     
         3 . The method of  claim 1  further comprising adding a sample to said reaction composition. 
     
     
         4 . The method of  claim 1  further comprising providing a detectably labeled probe or an intercalating dye. 
     
     
         5 . The method of  claim 1  further comprising thermocycling said reaction composition. 
     
     
         6 . The method of  claim 1  further comprising measuring a fluorescence emission of said reaction composition. 
     
     
         7 . The method of  claim 16  further comprising storing a realtime PCR reagent comprising said 0.0001 to 0.01% (w/w) MIT and at least one of said polymerase, said first primer, said second primer, and/or said nucleotides for at least 1 day. 
     
     
         8 . The method of  claim 7  comprising storing said realtime PCR reagent for at least 1 week. 
     
     
         9 . The method of  claim 7  comprising storing said realtime PCR reagent for at least 1 month. 
     
     
         10 . The method of  claim 7  wherein said realtime PCR reagent is used to prepare said reaction composition. 
     
     
         11 . A method of producing a realtime PCR reagent comprising mixing a PCR enhancer wherein said PCR enhancer is 0.0001 to 0.01% (w/w) MIT with at least one of a polymerase, a first primer, a second primer, and/or nucleotides. 
     
     
         12 . The method of  claim 11  wherein said realtime PCR reagent has a volume greater than 1 liter. 
     
     
         13 . The method of  claim 12  further comprising storing said realtime PCR reagent for at least 1 day. 
     
     
         14 . The method of  claim 12  further comprising storing said realtime PCR reagent for at least 1 week. 
     
     
         15 . The method of  claim 12  further comprising storing said realtime PCR reagent for at least 1 month. 
     
     
         16 . The method of  claim 12  further comprising mixing into said realtime PCR reagent a reference dye, a detectably labeled probe, an intercalating dye, a third primer, and/or a fourth primer.