Described herein are methods, compositions, and kits for library quantitation and qualification. Some embodiments relate to a method of library quantitation. For example, the method may include providing DNA fragments, amplifying the DNA fragments by polymerase chain reaction (PCR) in the presence of primers each labeled with a fluorophore. In these instances, only a predetermined number of fluorophores are attached to each DNA fragment. The method may further include detecting a fluorescent signal produced by the amplified DNA fragments and calculating a number of the amplified DNA fragments based on the detected fluorescent signal.
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