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View additional product information for NanoDrop™ 8000 Spectrophotometer - FAQs (ND-8000-GL)
43 product FAQs found
A report is shipped back with each NanoDrop instrument listing the services that were performed and any parts that were replaced. We also provide you with an Instrument Certification Sheet which shows that the instrument passed recalibration and recertification.
If you specifically request “as found” and “after service” data when setting up the NanoDrop instrument service, we can provide results of a Performance Verification before and after the servicing.
Upon receipt of confirmation of payment method, we will provide you a FedEx return shipping label and shipping instructions. Alternatively, we will waive the shipping cost if you decide to use your own carrier.
The NanoDrop 2000/2000c, NanoDrop 8000, NanoDrop Eight, NanoDrop Lite, NanoDrop Lite Plus, and NanoDrop One/OneC instruments all need to be shipped in their original shipping box and packing foam. If you do not have those materials, we will send you a box and packing foam at no cost. The NanoDrop 3300 instrument can be shipped in any box you may have available.
The cost for NanoDrop instrument service does not include any parts. If a part replacement is needed, we will contact you to obtain your authorization to replace the part.
You can pay for NanoDrop instrument service with a purchase order or credit card. We accept Master Card, Visa, or American Express.
The turnaround time for servicing NanoDrop instruments in the United States is 48 to 72 hours. The turnaround time in Canada is approximately 2 days longer, depending upon the time spent in Customs in either direction. FedEx weather-related delays can also extend the turnaround time.
We do not have onsite technicians to perform service for Thermo Scientific NanoDrop instruments located in the United States and Canada. All service for Thermo Scientific NanoDrop instruments is performed by our trained service technicians at our service depot in Madison, WI.
For NanoDrop instrument service in the United States and Canada, call 877-724-7690 (toll-free) and select option 4 for Technical Support or email nanodrop@thermofisher.com. For international service, please contact your local NanoDrop distributor.
All service for Thermo Scientific NanoDrop instruments is performed by our trained service technicians at our service depot in Madison, WI.
Visit this website (https://www.thermofisher.com/us/en/home/industrial/spectroscopy-elemental-isotope-analysis/molecular-spectroscopy/ultraviolet-visible-visible-spectrophotometry-uv-vis-vis/uv-vis-vis-instruments/nanodrop-microvolume-spectrophotometers/nanodrop-protein-quantification.html) for useful guidance on protein assays for NanoDrop One, NanoDrop Eight, and NanoDrop Lite spectrophotometers. View this guide (https://tools.thermofisher.com/content/sfs/manuals/T032-NanoDrop-2000-2000c-Protein-Measurements.pdf) for protein assays for NanoDrop 2000 spectrophotometer. View this guide https://tools.thermofisher.com/content/sfs/brochures/T010-ND-1000-&-ND-8000-Protein-Measurements.pdf) for NanoDrop 8000 spectrophotometer. Colorimetric assays such as the Pierce 660 nm, BCA, Bradford and Lowry, are generally used for uncharacterized protein solutions and cell lysates. Proteins that contain Trp, Tyr residues, or Cys-Cys disulphide bonds will absorb in the UV range (i.e., 280 nm) making absorbance spectroscopy a fast, convenient method for the quantitation of purified protein preparations using the Protein A280 application module.
Beer-Lambert Equation is as follows:
A = E * b * c
- A is the absorbance of the sample
- E is the wavelength-dependent molar absorptivity coefficient (or extinction coefficient) expressed in units of L/mol-cm
- b is the pathlength in cm
- c is the analyte concentration in mol/L or molarity (M).
For Nucleic Acid calculations, the Beer-Lambert equation is modified to use an extinction coefficient with units of ng-cm/mL. Using this extinction coefficient gives a manipulated equation:
c = (A * e)/b
- c is the nucleic acid concentration in ng/µL
- A is the absorbance of the sample
- e is the nucleic acid specific concentration factor in ng-cm/µL (dsDNA = 50 ng/µL; ssDNA = 33 ng/µL; RNA = 40 ng/µL; dsRNA = 40 ng/µL)
- b is the pathlength in cm
NanoDrop spectrophotometers use light pathlengths from 1.0 mm to as low as 0.05 mm (model dependent) to make measurements, allowing the quantification of samples up to 50 or 200 times more concentrated than traditional cuvette based spectrophotometers are able to measure. Applications such as Nucleic Acid and Protein A280 will display this absorbance as a 10 mm equivalent absorbance.
The sample concentrations are calculated according to Beer's Law which is volume independent. Beer's Law relates absorbance to concentration using a wavelength specific molar extinction coefficient or the use of a nucleic acid concentration factor and a pathlength of 1 cm.
The software is programmed to determine the best pathlength for the measurement based upon the absorbance signal of the sample at the analysis wavelength for the Nucleic Acids module, Protein A280 module, and Custom methods. On the other hand, the autopathlength function in the UV-Vis module monitors the entire wavelength range to optimize pathlength selection. The absorbance signal is normalized to a either 10 mm or 1.0 mm pathlength equivalent depending on module.
UV-Vis data is reported at a normalized 1.0 mm pathlength. As a result, absorbance values reported here will be reported 10-fold lower than those reported by the Nucleic Acid or Protein A280 applications.
Selecting autopathlength should be considered if the peak of interest is greater than 1.25 A at a 1.0 mm pathlength or to ensure that the entire sample absorbance spectrum remains within the linear response of the detector. With autopathlength deselected, the instrument uses only a 1.0 mm pathlength which is optimized for absorbance signal less than 1.25 A.
Both assays are mix-and-measure (minimal incubation times and no heating steps). Pierce 660 Assay has a more stable end-point. Pierce 660 reaction will not continue to progress and form aggregates.
When utilizing the measurement pedestal, NanoDrop spectrophotometers will report the Pierce 660 assay absorbance using a 1.0 mm pathlength. As a result, the absorbance displayed will be 10-fold lower than if measured in a conventional 10 mm cuvette. This reduction in signal does not compromise the sensitivity or linear range of the assay.
The Lowry module reports absorbance signal at a 1.0 mm pathlength rather than the conventional 10 mm pathlength resulting in a 10-fold reduction in absorbance signal. Consequently, the reduction in signal will not compromise the sensitivity or dynamic range of the assay. In the cases of extreme low signal, it is typically due to the formulation of the Lowry reagent from the specific vendors. We have had the greatest success with the Pierce product line of colorimetric assays.
General Information:
Wavelength (modified Lowry protocol for a NanoDrop spectrophotometer) is measured at 650 nm and normalized at 405 nm
Dynamic range of assay is 0.2 mg/mL to 4.0 mg/mL (BSA)
A poorly mixed Bradford assay will not properly complete and may form a gradient in the tube. This can affect reproducibility.
Some Bradford assay manufacturers' formulations will not provide sufficient absorbance to obtain an accurate and reproducible standard curve.
The Bradford reagent will form particulates as a reaction progresses. These particulates may cause light scatter during a sample measurement, resulting in higher than expected variation between measurements. Particulates can be minimized by limiting the incubation time to 10 min prior to measuring as per the Thermo Scientific Pierce protocol. In addition, a sample volume of 2 µL should be used for all colorimetric assay measurements.
To ensure accurate results, sample heating must be uniform for both standards and unknown samples.
When utilizing the measurement pedestal, NanoDrop spectrophotometers will report the BCA assay absorbance using a 1.0 mm pathlength. As a result, the absorbance displayed will be 10-fold lower than if measured in a conventional 10 mm cuvette. This reduction in signal does not compromise the sensitivity or linear range of the assay.
A purified protein should show a 260/280 purity ratio of approximately 0.63.
RIPA buffer produces a particularly strong absorbance signal at the 280 nm wavelength. As a result, it will either over estimate or under estimate protein concentrations and interfere with the protein purity ratio.
Protein samples in RIPA buffer should be quantified via the Pierce Protein 660 or BCA colorimetric assays using a full spectrum NanoDrop model.
Find additional tips, troubleshooting help, and resources within ourProtein Purification and Isolation Support Center.
The difference between the E1% and E0.1% values are the associated units for the results.
The E1% value has units of a 1% solution or 1 g/100mL. In the NanoDrop software this value is converted to an E0.1% value to display protein concentrations in mg/mL units.
Please refer to Tech Tip #6: Extinction Coefficients for further detail: https://assets.thermofisher.com/TFS-Assets/LSG/Application-Notes/TR0006-Extinction-coefficients.pdf
Protein concentrations obtained from the Protein A280 module are calculated using a 1% solution extinction coefficient (E1%) in place of the molar extinction coefficient. Each purified protein will have a unique E1% value which can be obtained from either references or, if produced commercially, from the vendor. The user may choose to use a known E1% value or input the molar extinction coefficient and molecular weight of the protein into the NanoDrop software to obtain the mg/mL protein concentrations.
If only a rough estimation is desired, the 1 Abs = 1 mg/mL can be used instead of a specific E1% value.
A conditional pass indicates that the calibration check % error for the 1.0 mm pathlength was >3% but = 5%.
More information about all of the calibration check tolerances may be found within the Calibration Check application.
Please review the following steps to take when/if your NanoDrop 8000 Spectrophotometer fails a calibration check:
- Clean the pedestal with the NanoDrop PR-1 Reconditioning Kit
- Repeat calibration check using a FRESH vial of NanoDrop CF-1 Calibration Fluid and FRESH blanking solution
- If instrument continues to be out of specification, Contact Technical Support
The Calibration Check software is included in the operating software under the Tools & Configuration tab. Select the Utilities & Diagnostics Application and the Calibration Check option.
All US & Canada instrument calibrations are performed in our Madison, Wisconsin service center by trained NanoDrop service technicians. There is a 48 hr turnaround time for all domestic service. International customers should contact their local distributors for service. The NanoDrop 8000 Spectrophotometer must be shipped in the original box and packaging. Please contact Technical Support or your local distributor if you require a box for shipment.
We recommend you check the calibration of your NanoDrop 8000 Spectrophotometer every 6 months using the NanoDrop CF-8 Calibration Check Kit.
You can easily check the calibration of your NanoDrop 8000 Spectrophotometer by using the NanoDrop CF-8 Calibration Check Kit, which contains two CF-1 Ampules and two 8-well PCR tube strips. The CF-8 kit is available from the Thermo Fisher website (Cat. No. CHEM-CF-8).
If re-calibration is ever needed, it is performed by a trained service technician. Due to the precise nature of the NanoDrop instrument design, these spectrophotometers rarely require re-calibration.
Pedestal surfaces may become "un-conditioned" and possibly inhibit proper liquid column formation required for measurement precision. Refer to this Cleaning and Reconditioning document for more information: http://tools.thermofisher.com/content/sfs/brochures/T005-NanoDrop%201000-&-NanoDrop%208000-Cleaning-and-Reconditioning.pdf
Measuring at or below the detection limit may result in SD values outside the precision specification. Refer to instrument user manual for detection limits for specific applications.
Evaporation can occur when samples remain on the pedestal beyond the time required to make one measurement, therefore we recommend always using a fresh aliquot for each measurement.
Poor sample homogeneity is commonly associated with non-homogeneous DNA samples. Due to the small sample volumes used for pedestal measurements it is important to mix the sample thoroughly. Samples containing large molecules such as genomic or lambda DNA are particularly susceptible to this phenomenon. If compatible with the protocol being used, heat the DNA samples to 63 degrees C and lightly finger vortex before measurement to ensure the nucleic acid is properly in solution.
NanoDrop CF-1 Calibration Fluid is the only photometric standard available to assess the accuracy of the NanoDrop spectrophotometer pedestal. If the control sample shows unexpected results the user should confirm the calibration of the instrument with the CF-1 fluid.
Some control examples include:
- Nucleic Acids: Fermentas GeneRuler Express DNA Ladder 100-5000 bp (Cat. No. SM1551)
- Protein: Pierce Bovine Serum Albumin Standard Ampules, 2 mg/mL (Cat. No. 23209)
The accuracy of NanoDrop spectrophotometers can be assessed by using NanoDrop CF-1 Calibration Fluid in conjunction with the calibration check application in the NanoDrop software.
Please review the following information regarding acceptable purity ratios, when using NanoDrop spectrophotometers:
- Nucleic acid samples: A260/A280 ratios of 1.8-2.2; A260/A230 ratios of 1.8-2.2 are generally considered pure
- Protein samples: A260/A280 ratio of approximately 0.63 is generally considered pure
Please review the following possible causes for 260/280 purity ratios to fall outside of the generally accepted range when using a NanoDrop spectrophotometer:
- The starting sample or the extraction method may result in contaminants in the final extract, e.g. guanidine (often from column based kits), phenol carryover or carbohydrates from the original cells (often a problem with plant samples). This is often best diagnosed by looking at the spectra, as many contaminants have characteristic profiles.
- Improper Blank: An incorrect or highly absorbent blanking solution or sample residue on the measurement pedestal may affect purity ratios.
- Sample Concentration: If the sample is nearing the lower detection limit of the instrument. Contributions from instrument noise may affect the shape of the spectrum. Check instrument specifications for lower detection limits. Purity ratios cannot be accurately calculated for samples close to the lower detection limits of the instrument. If a NanoDrop 2000c is being used, a cuvette may be used for samples at very low concentrations.