Pushing the Boundaries of UV LED

Real-time Analysis of Nucleic Acid Purity

Written by Stacy Hoge | May 6, 2020 6:16:46 PM

Conclusion
The KeyView Prep™ system allows for real-time 260/280 analysis that is both efficient and accurate.

Introduction

Since its conception in the 1940s, the 260/280 assessment of nucleic acid purity has routinely relied on spectrophotometric analysis and well-designed blanks. When implemented properly these analyses provide adequate quantity and quality assessments that inform downstream applications. Often, however, inherent biases in these protocols have misrepresented sample purity and negatively impacted downstream efforts. Two key biases seen in current 260/280 assessments are control mismatches and equipment differences. If the control solution does not closely match the sample in both ionic strength and pH the resulting over- or under-representation of purity undermines any subsequent analysis of that sample. Minor difference in wavelength (particularly at 280nm) can similarly invalidate further testing. Significant ratio differences have been seen even when comparing spectrophotometers that all lay within the 1nm wavelength specification. An issue germane to both intra- and inter-laboratory comparisons of the same sample. Here we report a novel, realtime 260/280 assessment of nucleic acid purity that is aimed at minimizing these inherent biases.

Real-time analysis of Nucleic Acid Purity