Multi-primer amplification method for tagging of target nucleic acids

Abstract

In certain embodiments, the present invention provides amplification methods in which nucleotide tag(s) and, optionally, a barcode nucleotide sequence are added to target nucleotide sequences. In other embodiments, the present invention provides a microfluidic device that includes a plurality of first input lines and a plurality of second input lines. The microfluidic device also includes a plurality of sets of first chambers and a plurality of sets of second chambers. Each set of first chambers is in fluid communication with one of the plurality of first input lines. Each set of second chambers is in fluid communication with one of the plurality of second input lines. The microfluidic device further includes a plurality of first pump elements in fluid communication with a first portion of the plurality of second input lines and a plurality of second pump elements in fluid communication with a second portion of the plurality of second input lines.

1 - 81 . (canceled) 
     
     
         82 . A method of preparing reaction products, the method comprising:
 providing M samples;   providing N assays;   mixing the M samples and N assays to form M×N pairwise combinations, each of the M×N pairwise combinations being contained in a closed volume;   forming M×N reaction products from the M×N pairwise combinations; and   recovering the M×N reaction products.   
     
     
         83 . The method of  claim 82  wherein the M samples are contained in M sets of first chambers and the N assays are contained in N sets of second chambers. 
     
     
         84 . The method of  claim 83  wherein the first chambers containing one of the M samples are associated with a set of second chambers, each of the set of second chambers containing one of the N assays. 
     
     
         85 . The method of  claim 84  wherein mixing the M samples and N assays comprises opening fluid lines to provide for fluid communication between the firsrt chambers containing one of the M samples and each of the set of second chambers containing one of the N assays. 
     
     
         86 . The method of  claim 82  wherein the first chambers are characterized by a volume of less than or equal to 100 nl. 
     
     
         87 . The method of  claim 86  wherein the volume is less than or equal to 40 nl. 
     
     
         88 . The method of  claim 82  wherein the second chambers are characterized by a volume of less than or equal to 10 nl. 
     
     
         89 . The method of  claim 88  wherein the volume is less than or equal to 2 nl. 
     
     
         90 . The method of  claim 82  wherein the closed volume is characterized by a volume of less than or equal to 100 nl. 
     
     
         91 . The method of  claim 82  further comprising thermocycling the M×N pairwise combinations. 
     
     
         92 . The method of  claim 82  wherein the M samples are provided at M sample ports of a microfluidic device. 
     
     
         93 . The method of  claim 91  wherein the M×N samples are recovered at the M sample ports of the microfluidic device. 
     
     
         94 . The method of  claim 82  wherein forming the M×N pairwise combinations is performed concurrently. 
     
     
         95 . The method of  claim 82  wherein forming the M×N pairwise combinations is performed sequentially.