| 主权项 |
1. A method of quantifying multiple target nucleic acid sequences in a sample, containing at least a first target nucleic acid sequence and a second target nucleic acid sequence, the method comprising:
(a) generating from a sample at least a plurality of first template molecules and at least a plurality of second template molecules, wherein said first template molecules comprise a sequence of a first target nucleic acid sequence and a first identity sequence (IS) tag, and said second template molecules comprise a sequence of a second target nucleic acid sequence and a second IS tag, and wherein said first IS tag comprises a first identification (ID) code and said second IS tag comprises a second ID code; (b) randomly distribute at least part of said first and at least part of said second template molecules generated from step (a) into individual reaction sites on a surface wherein the total number of said individual reaction sites and reaction volume are known, and the average number of said template molecules in each said individual reaction site is less than one; (c) generating at least one cluster of nucleic acid amplicons of said first template molecule and at least one cluster of nucleic acid amplicons of said second template molecules by clonal amplification or replication in spatially isolated said individual reaction sites on said surface; (d) simultaneously identifying said ID codes of all said nucleic acid amplicon clusters on said surface, wherein a unique ID code from any said nucleic acid amplicon cluster represents a positive identification of a target nucleic acid sequence in said sample; and (e) determining the quantity of at least said first and second target nucleic acid sequences in said sample by statistical analysis of respective positive numbers of identified unique ID codes of said first and second target nucleic acid sequences among said known number of individual reaction sites and said known reaction volumes on said surface. |
