Mineral hot start PCR method

Abstract

The present invention provides a method of performing hot start PCR reactions. The method is based on sequestration of magnesium ions in the form of a precipitate which renders a DNA polymerase inactive until the appropriate time in the PCR reaction when a certain temperature is reached and the magnesium ions are released from the precipitate. Also provided are kits comprising reagents and instructions for amplifying a target nucleic acid by the magnesium precipitate hot start.

We claim:  
     
         1 . A kit for amplifying a target nucleic acid, said kit comprising: 
 a. a container comprising a source of phosphate ions and a container comprising a source of magnesium ions, wherein combining the source of magnesium ions and the source of phosphate ions form a precipitate at a temperature below 34° C.; and    b. instructions for using the source of phosphate ions and the source of magnesium ions in amplification of the target nucleic acid.    
     
     
         2 . The kit of  claim 1  wherein the source of phosphate ions is phosphoric acid and the source of magnesium ions is magnesium chloride.  
     
     
         3 . The kit of  claim 2  wherein said kit further comprises a container comprising a DNA polymerase.  
     
     
         4 . The kit of  claim 3  wherein the DNA polymerase is TaqLA, Klentaq1, KlentaqLA, Pfu, Deep Vent or Tth.  
     
     
         5 . The kit of  claim 4  wherein said kit further comprises a container comprising deoxyribonucleoside triphosphates.  
     
     
         6 . The kit of  claim 2  wherein said kit further comprises a container comprising a mixture of at least two DNA polymerases selected from the group consisting of TaqLA, Klentaq1, KlentaqLA, Pfu, Deep Vent and Tth.  
     
     
         7 . The kit of  claim 6  wherein said kit further comprises a container comprising deoxyribonucleoside triphosphates.  
     
     
         8 . A kit for amplifying a target nucleic acid, said kit comprising: 
 a. a container comprising a reagent comprising a precipitate comprising a magnesium salt; and    b. instructions for using the precipitate to amplify the target nucleic acid.    
     
     
         9 . The kit of  claim 8  wherein said kit further comprises a DNA polymerase.  
     
     
         10 . The kit of  claim 9  wherein said DNA polymerase is TaqLA, Klentaq1, KlentaqLA, Pfu, Deep Vent or Tth.  
     
     
         11 . The kit of  claim 9  wherein said kit further comprises a container comprising deoxyribonucleoside triphosphates.  
     
     
         12 . The kit of  claim 8  wherein said kit further comprises a container comprising a mixture of at least two DNA polymerases selected from the group consisting of TaqLA, Klentaq1, KlentaqLA, Pfu, Deep Vent and Tth.  
     
     
         13 . The kit of  claim 12  wherein said kit further comprises a container comprising deoxyribonucleoside triphosphates.  
     
     
         14 . A kit for amplifying a target nucleic acid, said kit comprising instructions for using a source of phosphate ions and a source of magnesium ions in amplification of a target nucleic acid.