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
      • FTIR Accessories
  • Service
    • Service and Support Plans
    • Service Request Form
  • Applications
  • KnowledgeBase
  • Learning Center
    • Best Practice
      • 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
      • Training Registration Form
    • Webinars
    • eBooks
    • Theory
      • Theory of Molecular Spectroscopy
      • Chromatography
  • About Us
    • President’s Message
    • Contact
    • History
    • Careers
  • News
  • Events
  • Worldwide
  • Shop
  • My Account
  • Contact Us

Home / Applications / Measurement of Micro-Volumes of Proteins and Nucleic Acid

  • Industry

  • Technique

Measurement of Micro-Volumes of Proteins and Nucleic Acid

By Heather Haffner

PDF IconDownload This Application

January 5, 2024

Introduction

V-630BIO UV-Visible Spectrophotometer
V-630BIO UV-Visible Spectrophotometer

The SAH-769 One Drop accessory is used to measure micro-volumes of proteins and nucleic acids, either 0.6μL or 5 μL of sample with a 1 mm or 0.2 mm optical pathlength, respectively. The precise optical pathlength is controlled by covering the sample with a glass cover integrated into the cell unit. The cell and glass cover can be cleaned, simply by wiping with a laboratory tissue. This simple system can be used to measure samples quickly and easily. The shorter optical pathlength allows for measurements of high concentration samples without further dilution.

The V-630 Bio is double-beam spectrophotometer for high precision and accurate sample measurements, and can be operated using the cross-platform Spectra Manager or intelligent Remote Module (iRM) with color touch sensitive display. In either case, the Bio software includes standard programs for life science analyses. The Protein/Nucleic Acid program calculates either the protein or nucleic acid sample concentration from the ratio of 260/280nm absorbance. The Temperature Control program with optional Peltier thermostatted cell holder can be used for DNA thermal melting experiments.

This application note demonstrates the use of the one drop cell accessory with the V-630BIO system to measure small volume DNA samples.

Experimental

Measurement Conditions
Scan Speed200 nm/minData Interval0.5 nm
Bandwidth1.5 nmResponseMedium

Solutions of calf thymus DNA in KH2PO4/NaOH buffer (pH 7) at several concentrations were measured by using cells with 1 and 0.2 mm pathlengths. The measurement procedure for the SAH-760 One Drop accessory is shown in Figure 1.

 Measurement procedure using the SAH-769 One Drop Cell
Figure 1. Measurement procedure using the SAH-769 One Drop Cell.

Keywords

210-UV-0012, V-630BIO, UV-Visible/NIR, Life Science, Biochemistry, SAH-769 One Drop, Microsampling, nucleic acid, DNA, RNA, Protein

Results

The absorption spectra of various concentrations of calf thymus DNA using 1 and 0.2 mm pathlengths are shown in Figures 2 and 3, respectively.

Absorption spectra of calf thymus DNA using a 1 mm pathlength cell
Figure 2. Absorption spectra of calf thymus DNA
using a 1 mm pathlength cell.
Absorption spectra of calf thymus DNA using a 0.2 mm pathlength cell
Figure 3. Absorption spectra of calf thymus DNA using a 0.2 mm pathlength cell.

The corresponding calibration curves using the absorbance maxima at 260 nm are shown in Figures 4 and 5. Both curves demonstrate good linearity.

 DNA absorbance calibration curve in a 1 mm pathlength cell
Figure 4. DNA absorbance calibration curve in a 1 mm pathlength cell.
DNA absorbance calibration curve in a 0.2 mm pathlength cell
Figure 5. DNA absorbance calibration curve in a 0.2 mm pathlength cell.

The disc cell and cover glass were wiped clean after measuring the absorbance of the highest concentrated sample in the 1 mm pathlength (5 μL volume) cell. A 5μL solvent was then measured to evaluation sample cross-contamination of the disc cell. The DNA sample absorbance was 1.8832 while the solvent absorbance was 0.0002, indicating that wiping is enough to wash the sample from the cell.

This document has been prepared based on information available at the time of publication and is subject to revision without notice. Although the contents are checked with the utmost care, we do not guarantee their accuracy or completeness. JASCO Corporation assumes no responsibility or liability for any loss or damage incurred as a result of the use of any information contained in this document. Copyright and other intellectual property rights in this document remain the property of JASCO Corporation. Please do not attempt to copy, modify, redistribute, or sell etc. in whole or in part without prior written permission.

Featured Products:

  • 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

JASCO Application Note

Measurement of Micro-Volumes of Proteins and Nucleic Acid

Introduction

V-630BIO UV-Visible Spectrophotometer
V-630BIO UV-Visible Spectrophotometer

The SAH-769 One Drop accessory is used to measure micro-volumes of proteins and nucleic acids, either 0.6μL or 5 μL of sample with a 1 mm or 0.2 mm optical pathlength, respectively. The precise optical pathlength is controlled by covering the sample with a glass cover integrated into the cell unit. The cell and glass cover can be cleaned, simply by wiping with a laboratory tissue. This simple system can be used to measure samples quickly and easily. The shorter optical pathlength allows for measurements of high concentration samples without further dilution.

The V-630 Bio is double-beam spectrophotometer for high precision and accurate sample measurements, and can be operated using the cross-platform Spectra Manager or intelligent Remote Module (iRM) with color touch sensitive display. In either case, the Bio software includes standard programs for life science analyses. The Protein/Nucleic Acid program calculates either the protein or nucleic acid sample concentration from the ratio of 260/280nm absorbance. The Temperature Control program with optional Peltier thermostatted cell holder can be used for DNA thermal melting experiments.

This application note demonstrates the use of the one drop cell accessory with the V-630BIO system to measure small volume DNA samples.

Experimental

Measurement Conditions
Scan Speed200 nm/minData Interval0.5 nm
Bandwidth1.5 nmResponseMedium

Solutions of calf thymus DNA in KH2PO4/NaOH buffer (pH 7) at several concentrations were measured by using cells with 1 and 0.2 mm pathlengths. The measurement procedure for the SAH-760 One Drop accessory is shown in Figure 1.

 Measurement procedure using the SAH-769 One Drop Cell
Figure 1. Measurement procedure using the SAH-769 One Drop Cell.

Results

The absorption spectra of various concentrations of calf thymus DNA using 1 and 0.2 mm pathlengths are shown in Figures 2 and 3, respectively.

Absorption spectra of calf thymus DNA using a 1 mm pathlength cell
Figure 2. Absorption spectra of calf thymus DNA
using a 1 mm pathlength cell.
Absorption spectra of calf thymus DNA using a 0.2 mm pathlength cell
Figure 3. Absorption spectra of calf thymus DNA using a 0.2 mm pathlength cell.

The corresponding calibration curves using the absorbance maxima at 260 nm are shown in Figures 4 and 5. Both curves demonstrate good linearity.

 DNA absorbance calibration curve in a 1 mm pathlength cell
Figure 4. DNA absorbance calibration curve in a 1 mm pathlength cell.
DNA absorbance calibration curve in a 0.2 mm pathlength cell
Figure 5. DNA absorbance calibration curve in a 0.2 mm pathlength cell.

The disc cell and cover glass were wiped clean after measuring the absorbance of the highest concentrated sample in the 1 mm pathlength (5 μL volume) cell. A 5μL solvent was then measured to evaluation sample cross-contamination of the disc cell. The DNA sample absorbance was 1.8832 while the solvent absorbance was 0.0002, indicating that wiping is enough to wash the sample from the cell.

Keywords

210-UV-0012, V-630BIO, UV-Visible/NIR, Life Science, Biochemistry, SAH-769 One Drop, Microsampling, nucleic acid, DNA, RNA, Protein

This document has been prepared based on information available at the time of publication and is subject to revision without notice. Although the contents are checked with the utmost care, we do not guarantee their accuracy or completeness. JASCO Corporation assumes no responsibility or liability for any loss or damage incurred as a result of the use of any information contained in this document. Copyright and other intellectual property rights in this document remain the property of JASCO Corporation. Please do not attempt to copy, modify, redistribute, or sell etc. in whole or in part without prior written permission.
28600 Mary’s Court, Easton, MD 21601 USA • (800) 333-5272 • Fax: (410) 822-7526 • jascoinc.com/applications

Close

Designed in Tokyo. TRUSTED globally.

View our support plans

Connect with JASCO

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

Receive the latest promotions and special offers

  • This field is for validation purposes and should be left unchanged.
  • Careers
  • Press Kit
  • JASCO Privacy Policy
  • Sitemap
  • Environmental Policy

© , JASCO. All Rights Reserved.