Introduction
The DeNovix DS-11 Series Spectrophotometer / Fluorometer, equipped with SmartPath™ technology, enables accurate, reproducible measurements for both cuvette and microvolume measurements.
This document presents typical performance data comparing dsDNA measurements made using a DS-11+ versus measurements made using an Agilent 8453 Spectrophotometer across a concentration range measurable using a 1 cm cuvette.
Materials and Methods
A 250 ng/μL solution of fish sperm dsDNA (Amresco, cat #1b1509) was prepared in dH20. A series of eight dilutions ranging from ~0.5 to 75 ng/μL were then gravimetrically prepared to fall within the study concentration range.
Reference concentrations for the dilutions were determined using an Agilent 8453 (Agilent, Santa Clara CA) in a 10 mm quartz cuvette (Starna,#1-Q-10). The same cuvette was then used to measure six replicates of the same dilutions on the DS-11+ Spectrophotometer using the dsDNA EasyApps™ program and the Cuvette 10 mm mode.
Microvolume measurements were made using six 1 μL aliquots of dsDNA between 5 and 75 ng/μL. Fresh aliquots were used for each microvolume measurement. The sample solution was removed between each measurement by wiping the upper and lower sample surfaces with a clean, dry laboratory wipe.
Concentrations were calculated by applying Beer’s Law, using 10 mm equivalent 260 nm absorbance values.
Results
The linear response and accuracy correlation of the DS-11+ as compared to the reference Agilent 8453 Spectrophotometer is presented in Figure 1 and Table 1 below:
Agilent 1 cm cuvette n=6 | Agilent 1 cm cuvette n=6 | DS-11+ 1 cm Cuvette | DS-11+ 1 cm Cuvette | DS-11+ Microvolume | DS-11+ Microvolume |
---|---|---|---|---|---|
ng/μL | stdev | ng/μL | stdev | ng/μL | stdev |
0.53 | 0.01 | 0.58 | 0.02 | NA | - |
0.96 | 0.01 | 1.12 | 0.02 | NA | - |
2.5 | 0.02 | 2.73 | 0.02 | NA | - |
5.09 | 0.02 | 5.37 | 0.03 | 4.83 | 0.57 |
10.31 | 0.01 | 10.77 | 0.03 | 9.38 | 0.44 |
25.87 | 0.04 | 26.85 | 0.01 | 25.44 | 0.47 |
51.34 | 0.06 | 52.95 | 0.03 | 52.51 | 0.6 |
76.68 | 0.13 | 78.64 | 0.08 | 79.22 | 0.81 |
Summary
The DeNovix DS-11 Series exhibits outstanding linear and accuracy correlations to a calibrated Agilent 8453 Spectrophotometer. The dual cuvette and microvolume modes extend the concentration range capabilities (data not shown) and make the DS-11+ the instrument of choice for the broadest concentration range.
13-APR-2023