Skip to content

JASCO JASCO

  • News
  • Events
  • E-Store
  • My Account
  • Contact Us
  • Worldwide
Search
Click to view menu
MENUMENU
  • Products
    • Chromatography
      • HPLC
      • RHPLC
      • UHPLC
      • LC-MS
      • Preparative LC
      • Analytical SFC
      • Semi-Preparative SFC
      • Hybrid SFC
      • Fuel Analysis by SFC-FID
      • Preparative SFC
      • Supercritical Fluid Extraction
      • Chromatography Software
    • Molecular Spectroscopy
      • Circular Dichroism
      • High-Throughput CD
      • Vibrational CD
      • Circularly Polarized Luminescence (CPL)
      • Polarimeters
      • FTIR Spectrometers
      • FTIR Microscopy
      • FTIR Portable
      • Raman Microscopy
      • Palmtop Raman Spectrometer
      • Probe Raman
      • UV-Visible/NIR Spectrophotometers
      • UV-Visible/NIR Microscopy
      • Fluorescence Spectrophotometers
      • Film Thickness
      • Spectra Manager™ Suite
    • Refurbished
      • Refurbished HPLC Systems
      • HPLC Switching Valves
      • NRS-4100 Confocal Raman Microscopes
      • FTIR Accessories
  • Service
    • Service and Support Plans
    • Service Request Form
  • Applications
  • KnowledgeBase
  • Learning Center
    • Best Practices
      • Circular Dichroism Tips & Tricks for Biological Samples
      • CD Scale Calibration with ACS
      • Fluorescence Tips & Tricks
      • Raman Spectroscopy Tips & Tricks
    • Training Videos
      • ChromNAV
      • SF-NAV
      • Circular Dichroism
      • UV-Visible/NIR
      • Fluorescence
    • Training Seminars
      • Registration Form
    • Webinars
    • eBooks
    • Theory
      • Theory of Molecular Spectroscopy
      • Chromatography
  • About Us
    • President’s Message
    • Contact
    • History
    • Careers
  • News
  • Events
  • Worldwide
  • Shop
  • My Account

Home / Applications / Small Volume DNA Thermal Melting using a One Drop Accessory with Capillary Cell

  • Industry

  • Technique

Small Volume DNA Thermal Melting using a One Drop Accessory with Capillary Cell

By Leah Pandiscia, PhD

image_pdfView and Download PDF
August 15, 2022

Introduction

UV-Visible Spectrophotometer
V-630 UV-Visible Spectrophotometer

Thermal melting studies are commonly carried out in biochemical and biotechnology fields in order to obtain information regarding a biological sample’s thermodynamic properties. These properties can further elucidate the sample’s thermal stability and possible changes in structure and/or binding.

For many biological samples, it is much more convenient, cost effective, and efficient to use micro-volumes for structural and quantitative studies. Small volume DNA thermal melting analysis was made using the One Drop capillary jacket with a disposable quartz capillary cell, optical pathlength  0.5 mm and sample volume 3 µL.

This application note demonstrates the use of the one drop capillary jacket accessory for biological sample absorption measurements.

Experimental

Measurement Conditions
Measurement Wavelength260 nmData Acquisition Interval0.1°C
Temperature Gradient1°C/minResponseFast
Start Setting3 seconds +/-0.10°C

The measurement procedure is illustrated in Figure 1. The sample is drawn into the cell via capillary action and both edges of the capillary cell are sealed which helps to prevent volatilization of the sample. The capillary cell is then inserted into the capillary jacket and placed into a Peltier thermostatted cell holder. The jacket has a temperature sensor insertion port to ensure accurate measurements of the actual sample temperature.

Measurement procedure for filling the capillary cell
Figure 1. Measurement procedure for filling the capillary cell.

Keywords

UV-0008, V-630, UV-Visible/NIR, Microsampling, ETCS-761 Water-cooled Peltier thermostatted cell holder with stirrer, One Drop, Capillary jacket, Biochemistry, DNA thermal melting, Temperature ramping

Results

Figure 2 shows the result of measurement of Poly (dA-dT)-Poly(dA-dT). The absorption measurements were obtained in both the 10 mm rectangular cell (green) and the microsampling capillary cell (blue). The calculated melting temperature of Poly (dA-dT)-Poly(dA-dT) was 63.8ºC and 63.9ºC for the rectangular and capillary cells, respectively.

Thermal melting absorption data in a 10 mm rectangular cell (green) and capillary jacketed cell (blue)
Figure 2. Thermal melting absorption data in a 10 mm rectangular cell (green) and capillary jacketed cell (blue).

Conclusion

The results illustrate that the One Drop capillary cell and jacket are as reliable as the 10 mm rectangular cell for obtaining absorption measurements on extremely small volumes.

image_pdfView and Download PDF

Featured Products:

  • V-730 UV-Visible/NIR Spectrophotometer

    A general-purpose double-beam spectrophotometer with a compact space saving design

    V-730 UV-Visible Spectrophotometer

  • A high sensitivity UV-Visible/NIR Spectrophotometer with InGaAs detector for wavelengths up to1600nm

    V-780 UV-Visible/NIR Spectrophotometer

About the Author

Leah Pandiscia received her PhD from Drexel University where she studied Biophysical Chemistry. She is a Spectroscopy Applications Scientist.

JASCO provides services and support to a diverse range of industries throughout the world.

View our support plans

Connect with JASCO

  • Facebook
  • Twitter
  • LinkedIn
  • JASCO Sales
  • 800-333-5272

Receive the latest promotions and special offers

  • Careers
  • Press Kit
  • Privacy Policy
  • Sitemap

© , JASCO. All Rights Reserved.