Micro Volume Photometric Measurement of Nucleic Acid using TrayCell with UV-1280
Applications | 2015 | ShimadzuInstrumentation
Accurate quantification of nucleic acids in microvolumes is essential for molecular biology, diagnostics, and research where sample availability is limited. Traditional cuvettes require larger volumes and risk contamination, making ultra-micro volume approaches highly valuable.
This application note demonstrates microvolume photometric measurement of nucleic acids using the Shimadzu UV-1280 spectrophotometer equipped with Hellma TrayCell. The goal is to assess measurement repeatability and purity analysis of a 100 bp DNA ladder.
Samples of a Promega 100 bp DNA ladder (3 µl) were pipetted onto the TrayCell window. Baseline correction was performed using a reference blank. Measurements were conducted at 260 nm and 280 nm in DNA quantitation mode to determine concentration and purity.
Ten replicate measurements of the DNA ladder at 260 nm yielded an average absorbance of 0.275, a standard deviation of 0.004, and an RSD of 1.32%. Purity ratios (A260/A280 and A260/A230) were within expected ranges (1.8–1.9 and >2.0 respectively), confirming minimal contamination.
Advancements may include integration with automation platforms, expanded wavelength ranges for RNA and protein profiling, and coupling with microfluidic systems for high-throughput analysis.
The combination of Shimadzu UV-1280 bio method mode and Hellma TrayCell enables reliable microvolume nucleic acid measurements with excellent precision and purity assessment.
Shimadzu (Asia Pacific) Pte. Ltd. (2015). Application Note AD-0088: Micro Volume Photometric Measurement of Nucleic Acid using TrayCell with UV-1280.
UV–VIS spectrophotometry
IndustriesPharma & Biopharma
ManufacturerShimadzu
Summary
Importance of the Topic
Accurate quantification of nucleic acids in microvolumes is essential for molecular biology, diagnostics, and research where sample availability is limited. Traditional cuvettes require larger volumes and risk contamination, making ultra-micro volume approaches highly valuable.
Objectives and Study Overview
This application note demonstrates microvolume photometric measurement of nucleic acids using the Shimadzu UV-1280 spectrophotometer equipped with Hellma TrayCell. The goal is to assess measurement repeatability and purity analysis of a 100 bp DNA ladder.
Methodology
Samples of a Promega 100 bp DNA ladder (3 µl) were pipetted onto the TrayCell window. Baseline correction was performed using a reference blank. Measurements were conducted at 260 nm and 280 nm in DNA quantitation mode to determine concentration and purity.
Instrumentation Used
- Shimadzu UV-1280 UV-VIS spectrophotometer with six bio method mode protocols (DNA Quantitation, Lowry, BCA, CCB, Biuret, UV).
- Hellma TrayCell ultra-micro cell with 1 mm and 0.2 mm path length caps, enabling sample volumes of 0.7–10 µl and virtual dilution factors of 1:10 and 1:50.
Main Results and Discussion
Ten replicate measurements of the DNA ladder at 260 nm yielded an average absorbance of 0.275, a standard deviation of 0.004, and an RSD of 1.32%. Purity ratios (A260/A280 and A260/A230) were within expected ranges (1.8–1.9 and >2.0 respectively), confirming minimal contamination.
Benefits and Practical Applications
- High reproducibility with minimal sample volume and reduced dilution errors.
- Rapid analysis suitable for routine quality control in genomics, proteomics, and clinical laboratories.
- Cost and time savings by eliminating extensive cleaning steps and sample preparation.
Future Trends and Potential Applications
Advancements may include integration with automation platforms, expanded wavelength ranges for RNA and protein profiling, and coupling with microfluidic systems for high-throughput analysis.
Conclusion
The combination of Shimadzu UV-1280 bio method mode and Hellma TrayCell enables reliable microvolume nucleic acid measurements with excellent precision and purity assessment.
References
Shimadzu (Asia Pacific) Pte. Ltd. (2015). Application Note AD-0088: Micro Volume Photometric Measurement of Nucleic Acid using TrayCell with UV-1280.
Content was automatically generated from an orignal PDF document using AI and may contain inaccuracies.
Similar PDF
Accuracy in Microvolume Measurements. Investigating the Precision and Reproducibility of Cary 50, 300 and 4000 series Instruments for Microlitre samples
|Agilent Technologies|Applications
NOTICE: This document contains references to Varian. Please note that Varian, Inc. is now part of Agilent Technologies. For more information, go to www.agilent.com/chem. Sample Analysis Report Number 49 Accuracy in Microvolume Measurements. Investigating the Precision and Reproducibility of Cary…
Key words
dna, dnatraycell, traycellhellma, hellmapathlength, pathlengthlimits, limitsnucleic, nucleicspectrophotometer, spectrophotometeralignment, alignmentmicrovolume, microvolumemonochromator, monochromatorscan, scannormal, normalevaporative, evaporativedetection, detectionmonochromation
Quantitation of Nucleic Acids – Trace Measurement Using TrayCell and Nano Stick –
2018|Shimadzu|Applications
LAAN-A-UV-E072 Application News No. A577 Spectrophotometric Analysis Quantitation of Nucleic Acids – Trace Measurement Using TrayCell and Nano Stick – Ultraviolet-visible (UV-VIS) spectrophotometers are used in quantitative and qualitative analysis of substances in many fields. Purity confirmation and quantitation of…
Key words
nucleic, nucleicabsorbance, absorbancetraycell, traycellquantitation, quantitationstick, stickvalue, valuenano, nanomeasurement, measurementindicate, indicatedripped, drippedcorrelation, correlationacid, acidwavelength, wavelengthnews, newsdeviation
Shimadzu UV Talk letter Vol. 21
2023|Shimadzu|Others
C101-E176A Vol. 21 Effect of Measurement Parameters on UV-VIS Absorption Spectra ------- 02 Applications Examples of Analysis Using Micro-Volume Measurement Cells ------- 06 Q&A What is the difference between absorbance and optical density (O.D.)? ------- 13 PQY-01 Photoreaction Evaluation System…
Key words
traycell, traycellstick, sticklight, lightnano, nanomeasurement, measurementwavelength, wavelengthintervals, intervalswidth, widthslit, slitcells, cellsoptical, opticallightway, lightwaypath, pathabsorption, absorptionlength
Micro Volume Quantitation Measurement of Nucleic Acid using Nano Stick with UV-1280
2015|Shimadzu|Applications
Application News UV-1280, Nano Stick, Nucleic Acid Micro Volume Quantitation Measurement of Nucleic Acid using Nano Stick with UV-1280 AD-0097 Introduction An ultraviolet-visible (UV-VIS) spectrophotometer can be used for quantitation of nucleic acids and proteins and such samples are…
Key words
sinco, sincostick, sticknucleic, nucleicmicro, micronano, nanospectrophotometer, spectrophotometerdna, dnavolume, volumeabsorbance, absorbancenews, newspitch, pitchacids, acidsserially, seriallycuvette, cuvetteoverlapped
