Methods for forensic DNA quantitation

摘要

Described herein are methods and devices for nucleic acid quantification and, in particular, to microfluidic methods and devices for nucleic acid quantification. In certain embodiments methods of quantifying a target nucleic acid without the need for amplification are provided. The methods involve, in some embodiments, allowing a binding agent to become immobilized with respect to the target nucleic acid. In some cases, the binding agent comprises a signaling moiety that can be used to quantify the amount of target nucleic acid. In another aspect, the quantification can be carried out rapidly. For example, in certain embodiments, the quantification can be completed within 5 minutes. In yet another aspect, samples containing a low amount of target nucleic acid can be quantified. For instance, in some cases, samples containing less than 100 nanograms per microliter may be quantified. Also described are devices and kits for performing such methods, or the like.

主权项

1. A method for quantifying an unamplified double stranded target nucleic acid in a sample fluid containing or suspected of containing the unamplified double stranded target nucleic acid, comprising:
combining in a microfluidic channel located in a biochip, said biochip further comprising a plurality of microfluidic channels and active areas for sample manipulation, at least a portion of the sample fluid and a probe fluid containing a binding agent comprising a signaling moiety to form a test fluid; heating said test fluid to denature the unamplified double stranded target nucleic acid and cooling said test fluid to bind the binding agent to the denatured unamplified target nucleic acid; maintaining the test fluid stationary in the microfluidic channel at a detector region; detecting the signaling moiety in the test fluid at said detector region; and quantifying the denatured unamplified target nucleic acid in the test fluid based on the detected signaling moiety; wherein the denatured unamplified target nucleic acid in the test fluid is quantified within 1 minute of combining the portion of the sample fluid and said probe fluid.