Method of preparing a reaction mixture and related products

Abstract

The invention relates to a method of preparing a reaction mixture for Polymerase Chain Reaction (PCR) assay and a solution set for PCR. The method comprises providing a sample solution comprising a biological sample to be amplified in said PCR assay and first colorant providing the solution a first color, providing a reagent solution comprising at least one other substance required for performing said assay and second colorant providing the solution a second color different from the first color, and mixing the sample solution and the first reagent solution for providing a mixed solution to be subjected to the PCR process, the mixed solution having, due to said first and second colorants, a third color different from the first and second colors. The invention significantly aids in pipetting PCR assays.

1 - 34 . (canceled) 
     
     
         35 . A method of preparing a reaction mixture for Polymerase Chain Reaction (PCR) assay, comprising:
 (i) providing a first reagent solution comprising at least one substance for performing said assay,   (ii) providing a second reagent solution comprising at least one other substance for performing said assay,   (iii) mixing the first and second reagent solutions for providing a mixed solution to be subjected to the PCR process,   wherein the first reagent solution contains first colorant providing the first reagent solution a first color; the second reagent solution contains second colorant providing the solution a second color different from the first color; said mixing yields a mixed solution having, due to said first and second colorants, a third color different from the first and second colors; and   wherein the third color is detected visually.   
     
     
         36 . The method of  claim 35 , further comprising providing a third reagent solution comprising at least one further substance for performing said assay, the third reagent solution containing a third colorant providing the solution a fourth color different from the first, second and third colors; and
 mixing the third reagent solution with the first and second reagent solutions for providing a mixed reagent solution having, due to said first, second and third colorants, a fifth color different from the first, second, third and fourth colors.   
     
     
         37 . The method of  claim 35 , further comprising providing a third reagent solution containing third colorant providing the solution a fourth color different from the first, second and third colors; and
 individually mixing the first reagent solution with said second and third reagent solutions for obtaining first and second mixed solutions having third and fifth colors, respectively, different from each other and the first, second and fourth colors.   
     
     
         38 . The method of  claim 37 , wherein the first reagent solution is a PCR or qPCR master mix, the second reagent solution contains a first set of primers and the third reagent solution contains a second set of primers different from the first set of primers. 
     
     
         39 . The method of  claim 35 , wherein the first reagent solution comprises at least one substance for the PCR assay, which is a polymerase, a primer, a plurality of dNTPs, a fluorescent qPCR dye, or a probe. 
     
     
         40 . The method of  claim 39 , wherein the second reagent solution comprises at least one other substance not in the first reagent solution and for the PCR assay, which is a polymerase, a primer, a plurality of dNTPs, a fluorescent qPCR dye, or a probe. 
     
     
         41 . The method of  claim 40 , wherein the first reagent solution comprises a polymerase or a polymerase buffer; and the second reagent solution comprises at least one of primers, a probe, and a sample. 
     
     
         42 . The method of  claim 35 , wherein the reaction mixture is for use in a quantitative PCR assay. 
     
     
         43 . The method of  claim 42 , wherein the reaction mixture comprises a fluorescent agent, and wherein the absorbance peaks of the colorants do not overlap with the emission or excitation wavelength of the fluorescent agent or the total absorbances of the reaction mixture at the emission or excitation wavelengths of the fluorescent agent are less than 0.05 when measured with a 1 mm light path. 
     
     
         44 . The method of  claim 42 , wherein the mixed solution is translucent. 
     
     
         45 . The method of  claim 42 , wherein the mixed solution is transparent. 
     
     
         46 . The method of  claim 35 , wherein the first reagent solution or the second reagent solution further comprises a biological sample to be amplified in the PCR assay. 
     
     
         47 . The method of  claim 35 , wherein the first reagent solution or the second reagent solution is a PCR master mix, a qPCR master mix, a PCR premix or a qPCR premix. 
     
     
         48 . The method of  claim 35 , wherein the first reagent solution or the second reagent solution further comprises a sample elution buffer. 
     
     
         49 . The method of  claim 35 , wherein the first reagent solution or the second reagent solution further comprises a sample dilution buffer. 
     
     
         50 . The method of  claim 35 , wherein at least one of the first and second colorants is selected from Quinoline yellow, Xylene cyanol, Brilliant Blue, Patent Blue, Indigocarmine, Acid Red 1, m-Cresol Purple, Cresol Red, Neutral Red, Bromocresol Green, Acid Violet 5, Bromo phenol blue, and Orange G. 
     
     
         51 . The method of  claim 35 , wherein at least one of the first reagent solution or the second reagent solution is a concentrate. 
     
     
         52 . The method of  claim 35 , further comprising subjecting the mixed solution to a quantitative PCR process. 
     
     
         53 . The method of  claim 38 , further comprising subjecting the mixed solution to a quantitative PCR process. 
     
     
         54 . The method of  claim 35 , wherein the mixing is carried out by pipetting the reagent solutions to one or more wells of a microtiter strip or plate.