Frequently Asked Questions

• Can our Real-time PCR system run on a Mac?

  • No. The software is compatible with Windows 2000 and Windows XP.

• Can our Real-Time PCR system run on Windows Vista?

  • No.

• Can our Real-Time PCR system run on Windows Vista?

  • No

• What type of processor is required to run our Real Time system software?

  • The software will only run on a computer with an Intel Centrino processor. The software will not run on systems using a Celeron processor.  

• Can I use the efficiency for each individual sample to do quantification?

  • Theoretically yes. We recommend to use the average efficiency of all samples for each gene (the same primer set) to do the quantification.  In most cases, only if you want to know how much do tissue, or if treatment specific inhibitory and activatory effects of PCR reaction for the same gene exist in your experiment, you might want use the averaged efficiency for replicates instead for each gene to do the quantification computation. Even though, we recommend you use more replicates (more than 5 or 6) instead of only triplicates to run your Real-time PCR experiment.

• Is there a simple way to tell the data I submit is pure raw data, or baseline subtracted data

  •  Yes. You can have a quick look at the data you exported from the Real-time PCR, especially the first 15 rows. The number of pure raw data should always be larger than the one after baseline subtraction. If you find any negative values, that means you may have exported the baseline subtracted data. The instrument uses the minimal value as baseline. You need to choose the data set with a larger value.

• What is the difference between a gradient and a non-gradient system?

  • The difference is that a temperature gradient can be set in the gradient system to facilitate the determination of the correct annealing temperature.

• Is there a heated lid on the Swift and Spectrum cyclers?

  • Yes, The Swift Maxi and Spectrum 48 cyclers have an adjustable temperature heated lid. The heated lid default temperature is 105°C. C on the Swift Maxi and the Spectrum 48.

• What is the range to which the gradient can be set?

  • 30° on the Swift Maxi and 25°C on the Spectrum 48.

• What is the accuracy of the temperature in the blocks?

  • Swift Maxi: 0.5°C @ 95°C, 0.3°C @ 72°C, 0.2°C @ 55°C
  • Spectrum 48: =0.1°C

• Can the temperature in the Swift and Spectrum wells be measured with a standard lab thermometer?

  • Yes, but a digital thermometer is preferable.

• Do the Swift and Spectrum cyclers use Peltier for heating and cooling?

  • Yes. The Swift Maxi uses a 6 element Peltier and the Spectrum 48 uses a unique, 4 element, side mounted Peltier.

• Is it possible to add a 5th channel to Swift Spectrum RT 48 ?

  • Yes. This is available as an optional, factory fitted upgrade. 

• What type of block is utilized on the Spectrum 48?

  • A 48 well, linear block ( 2 rows of 24 ) with a unique, patented Peltier design that allows for exceptional temperature accuracy and fast heating and cooling ramp rates.

• What chemistries and consumables can be used in the Spectrum 48?

  • 100% open platform chemistry and consumables. Can accommodate all current RT chemistries, and dyes, except for Cy-5.

• Can the Spectrum 48 operate using any quality of reaction tube?

  • Yes, but to obtain the maximum signal intensity, only optical quality reaction vials should be used. The poorer the quality, the higher the loss of signal.

• What type of detection system is used in the Spectrum 48?

  • A broad band, PMT detector is used.

• How is the fluorescence activated?

  • Activation is by blue diode light emission.

• What are the advantages of a diode/PMT system?

  • Lower running costs, low light scatter, no need for regular alignment, low replacement costs.

• How do I obtain software upgrades?

  • Upgrades are free, through the internet.

• How is the Spectrum 48 interfaced with the computer?

  • The instrument is connected to the computer by a serial cable.

• Can I transfer protocols from other brands to the Swift and Spectrum 48 cyclers?

  • Yes. Both instruments have adjustable ramp rates allowing the transfer of protocols used on other brands.

• How do you program the gradient function on the Swift Maxi and Spectrum 48 cyclers?  

  • Swift Maxi: The software will automatically recognize the gradient block. The minimum and maximum temperatures are defined and the software will automatically calculate the correct gradient depending on the rage selected.
  • Spectrum 48: From the drop down menu select Gradient. Enter the gradient range in the relevant windows. The software automatically calculates the gradient depending on the range selected.

• How do you view the gradient on the Swift Maxi and Spectrum 48 Cyclers?

  • Swift Maxi: The gradient is continuously displayed in the main window.
  • Spectrum 48: The gradient is displayed in the gradient window from the Gradient drop down menu.

• Can I use the gradient function with the in-situ block?  

  • No. We recommend that you optimize the PCR in a tube before proceeding to the actual experiment.

• Are there different ways to start a Swift Maxi or Spectrum 48 program?  

  • No. Simply turn the instruments on and program through the arrow keys or through the Spectrum software.

• Can I use the Swift Maxi to amplify DNA for use with the ABI sequencing kits? 

  • Yes, but use the correct ramp rates as specified for the ABI kit.

• How long has Esco been making thermal cyclers? 

  • Esco has been in the thermal cycler business since 2006. . These models have been marketed all over the world.

• Can I purchase an extended warranty for my Swift and Spectrum cyclers?  

  • Yes. Extended warranties are available. Please contact our Customer Service department for more information and pricing.

• Which PCR tubes fit best into the blocks?  

  • All conventional tubes, from any manufacturer. We recommend the use of good quality, preferably optical grade tubes in our Spectrum 48 cycler. Poor quality tubes will result in loss of signal and lower sensitivity.
  • Our blocks accommodate 0.2 and 0.5ml tubes.

• Is it necessary to perform oil overlays when using our cyclers? 

  • No. The heated lid prevents the formation of condensation in all tube types used.

• What programming possibilities does the software offer?  

  • Our cyclers offer a high degree of programming flexibility. The software allows the variation of several critical parameters in PCR, including touchdown; touch-up, incremental changes in time extension, ramp-rate changes and incremental ramp-rate changes.  

• Why is the temperature not measured in the sample?  

  • A separate sample sensor can have several disadvantages, such as blocking a sample position. Obstruction caused by supply cables and possible control errors may occur with an incorrectly positioned sample probe. In addition, sample control probes usually use mineral oil that has different thermal properties from PCR buffers. Our software uses set algorithms to precisely control the sample temperature based on the size tube and sample volume used.

• What advantages does the gradient function have, and when can it be used?  

  • The gradient function can be considered an aid for optimizing PCR. Instead of repeatedly testing new annealing temperatures in several successive experiments, different temperatures can be tested in parallel during one run. Users can therefore optimize their PCR conditions in a much shorter time and with considerably less effort (and money!). This application is particularly advantageous in basic and clinical research.

• Is in situ PCR on slides possible?  

  • Yes. An in situ slide block can be purchased for the Swift Maxi.

• Are interchangeable blocks available?

  • The Swift Maxi has a choice of 8 blocks, including a gradient, in situ, 386 well and three dual blocks. There are no interchangeable blocks for the Spectrum 48 cycler.

• What is the function of the dual blocks?

  • The dual block allows the simultaneous running of two different PCR experiments. Each block is fully programmable and independent of the other block. It is like having two PCR instruments in one.

• Are there temperature overspills when both blocks are in use?

  • No. Each block is fully insulated.

• What is the Hot Start technique?  

  • The Hot Start technique is a method designed to reduce background in PCR. All reaction components, with the exception of Taq , are heated to the initial denaturation temperature. The Taq polymerase is then added, and thermal cycling is begun.

• Do the samples in the cycler have to be cooled down to 4°C after a PCR?  

  • Cooling down to 4°C is hardly ever necessary! After all, the DNA in the PCR experiment is heated 25 to 30 times up to almost 100°C. Therefore, there are no active DNases present that could degrade the DNA at room temperature.

•   How do Peltier elements work?  

  • A Peltier element functions like an electric heat pump. Heating and cooling are in response to the direction of the electrical current. As one side of a Peltier element is heated, the other side cools. Utilization of Peltier technology allows rapid heating and cooling of the thermal cycler block, therefore decreasing overall experiment times.

• What material is used in the blocks of the Swift Maxi and Spectrum 48 cyclers?  

  • The blocks are made from a special aluminum alloy. The choice of material was made to ensure the lowest possible heat capacity (thus enabling fast temperature ramping), as well as sufficient hardness for a long service life (no deformation of the wells through heat and pressure, nor warping of the block from constant use and thus long-term optimum heat transfer from the block to the reaction tube).

• What advantages are there for customers in connecting their Swift Maxi to a PC?  

  • The main advantage for users is the recording and storage of data during a PCR run.
  • Report generation and storing on a PC improve user overview, simplifying the generation of reports.
• I have to work with GLP (Good Laboratory Practice) and validate my Swift Maxi and Spectrum 48 accordingly. How can I do this?  
• For validation, Esco offers a temperature validation system with which users can test the Thermocycler themselves.

• Can Esco calibrate or validate a Swift cycler?

  • Yes. For this purpose, please contact ESCO.

• Can a printer and a PC be connected at the same time to a Swift Maxi?

  • No  

• What are the key benefits of the Swift Spectrum?

  • The Swift Spectrum brings advanced PCR technology to the bench top with its highly intuitive operation and analysis software, and its extremely robust, compact design. The Swift Spectrum combines the advanced, Peltier block design with high-speed gradient thermal block with exceptional temperature ramp rates and accuracy. The optical detection is sensitive and offers a wide dynamic range so you can analyze more samples with greater flexibility in a single run. The block can be fragmented allowing multiple sample groups to be analysed at the same time.

• Can I run my established protocols on the Spectrum 48 and Swift Maxi cyclers?

  • Yes, established protocols can easily be adapted to these cyclers. The temperature ramp rates can be fine-tuned to mimic the characteristics of other PCR platforms. Furthermore, the Swif t Maxi' s gradient capabilities enable you to test a range of temperatures across 12 columns of a single plate so that you can easily re-optimize your primer annealing temperatures if required, when switching from a slower ramping system to the higher speed.sytem.

•   How fast are the blocks?
  •  

    Maxi:
    	Heating	4°C/s
    	Cooling	3.5
    Spectrum 48	Heating	4.0
    	Cooling	3.5

• Is there an even excitation of the LED over the 48 positions? 

  • Yes. Because of the small block size, excitation is 100% even over the entire area of the block.

•   Can ROX be used for normalization?  

  • There is no need to use ROX for normalization.

• Does the Spectrum 48 require any specific chemistry?  

  • The Spectrum 48 is an open system that does not require or prefer any specific chemistry. All intercalating dyes or probe-based detection formats are supported.

• What kind of tubes can I use with the Spectrum 48?  

  • All standard tubes with optically clear flat sealing can be used.

• What is the excitation source of the Spectrum 48?

  • The Spectrum 48 x uses a 48 LED array with an excitation wavelength of 470 nm, which is able to effectively excite the most common dyes used in real-time PCR.

• Is the Spectrum 48 capable of multiplexing? 

  • Yes, the Spectrum 48 is capable of multiplexing. With the Spectrum 48 , duplex assays can be performed.

• How long does it take to detect all 48 samples?  

  • 24 seconds are required for data acquisition from 48 samples.

• Can I perform relative quantification with the Spectrum 48?  

  • Yes, a data analysis module for relative quantification (??C t method) is included in the software.
  • 33  What is the bandwidth for every filter?  

    520 nm	±10 nm
    550 nm	±10 nm
    580 nm	± 5 nm
    605 nm	±15 nm

• Is it possible to detect more than one dye per filter during a single run? 

  • No, it is not possible to define more than one dye per filter for a single run.

• To what standards of documentation do the Swift and Spectrum cyclers conform? 

  • GLP-compliant documentation of all, PCR , real-time PCR and device data.

• Is a SNP-analysis possible with the Spectrum 48? 

  • Yes, an analysis module to suit this application is a part of the software package.

• Can I use more than one dye for the analysis mode “Relative Quantification/ Monoplex”?

  • No.

• How do we prevent an interaction between the emission spectra of different dyes?

  • During color calibration at time of installation, the software determines a sensitivity code for every dye. The overlapping of two dyes can then be accounted for and will be subsequently avoided during analysis.

• What is the meaning and function of the gain setting for the dyes? 

  • For every dye and every filter the CPMs (an advanced type of photomultiplier) will adjusted to a defined sensitivity. This sensitivity is adjusted to fall within a defined range, and the specific data is stored for each scan.

• It is possible to run more than one Spectrum 48 on one computer? 

  • This is not possible due to requirements of data security and the proper assignment of customized calibration data.

• Can the software calculate the efficiency for every curve?

  • The software does not calculate the efficiency for each individual amplification curve. PCR efficiency is calculated by the software for each standard curve. To ensure the most comparative, reliable data, it is therefore necessary to optimise your PCR reactions to a comparable PCR efficiency as the standard curve.

• How often is it necessary to repeat the color calibration? 

  • After the first installation and each time a new dye will be used, it is necessary to perform a color calibration. This data is then stored in the database (not in the instrument). If you wish to evaluate the calibration data over time, it is possible for standard users to perform color calibrations.

• Could there be an effect on results when volumes differ across the wells of a calibration plate? 

  • In this case the fluorescent intensities are different. It is therefore possible that analysis results may be subsequently distorted. However, this influence is minimised due to Eppendorf's means of calibration, turning the plate to average effect the readings during the calibration.

• How long should the device be allowed to warm up, to enable a self-check of the service functions? 

  • The warm up time should be 15 minutes (between turning on the instrument and the software and checking the service functions). The optical module requires this time to reach a constant working temperature. The same is recommend for endpoint detection. Using a conventional PCR program the optimal temperature will be reached during the first cycles, where normally no signal increase will be detected.

• How can I test the cleanliness of the block? 

  • because of the block design and the way the emissions are read, there is no need to do this.

• What are the minimum specifications for the Spectrum 48 operating computer?

  • Intel Centrono/ Intel Pentium III @ 1.0 GHz, min. 512 MB RAM
  • 4 GB hard-disk storage (up to 1 GB required for software and database)
  • Serial Port
  • CD-ROM writer + software for writing CDs (integrated or external)
  • Oprating System: Windows2000, XP or higher

• Where is the product made?

  • The products are made in China according to Esco specifications.

• How did Esco enter the PCR market and what background does Esco have?

  • The decision to enter into this market is in accordance to Esco's product development strategy to offer top quality products to the scientific community.

• Why is the 48 well format better?

  • Research applications seldom require a larger capacity. The 48 well block is small enough to guarantee excellent temperature accuracy, spread and fast ramping rates.

• Why is our Real-Time Cycler better?

  • The unique block design, detection from the bottom of the tube, fast ramp rates and sensitivity.

• How about the CE mark?

  • The products carries the CE mark.

• How many channels does the instrument have?

  • This is a 4 channel instrument with three usable dye channels.

• Do we supply consumables?

  • We do not supply consumables, but the Spectrum 48 is an open platform instrument.