The World’s fastest Real-Time PCR Thermal Cycler User Manual This Manual Copyright © 2014 BJS Biotechnologies
Page |2 xxpress, xxplates, xxsealer, and their respective logos are registered trademarks of BJS Biotechnologies Ltd. or one of its affiliated companies in the UK and other countries. All rights reserved. No part of this manual may be reproduced, stored in a retrieval system, or transmitted, by any means or in any form, without the prior permission of BJS Biotechnologies Ltd (BJS Bio). The information contained in this manual is subject to change without notice.
Page |3 Contents 1 2 3 4 5 6 7 7.1 7.2 8 8.1 8.2 8.3 8.4 9 10 Glossary of Terms and Abbreviations ............................................................. 4 Symbols Used in this Instruction Manual ........................................................ 5 Unit Description ............................................................................................... 6 Technical Specification ....................................................................................
Page |4 1 Glossary of Terms and Abbreviations ANSI BJS Bio American National Standards Institute BJS Biotechnologies Limited, UK Company Number 02959160 Deepwell Plate Plate with an SBS footprint featuring 48, 96 or 384 wells with a larger volume than microplates Deepwell plate Electro-Magnetic Compatibility DWP EMC Microtitre Plate MTP PCR PPE Real-Time PCR SBS Plate with an SBS footprint featuring 24, 48, 96 or 384 wells Microtitre plate Polymerase Chain Reaction Personal Protective Equipment Poly
Page |5 2 Symbols Used in this Instruction Manual The following advisory symbols are used in this manual. Table 1: Advisory Symbol Meanings Indicates a Risk of Electric Shock which could, if not avoided, result in serve injury or death. DANGER Indicates a Burn Hazard which could, if not avoided, result in serve injury or death. DANGER Indicates a Risk of Explosion which could, if not avoided, result in serve injury or death.
Page |6 3 Unit Description The xxpress Real-Time Thermal cycler provides a safe and controlled method heating and cooling the test samples placed within the xxplates whilst measuring the level of fluorescence emitted by the samples. xxpress has five fluorescence measurement channels that enable fluorophores with a wide range of excitation and collection frequencies to be used.
Page |7 12 15 14 16 13 17 11 10 Table 2: Unit Features 1 2 3 4 5 6 7 8 9 10 xxpress Power On/Off Button Front Door Front Door Release Button xxplate location xxpress Display and Touch Screen Front USB Connector Rear Cover Air Intake Vents and Filters (under unit) Power on LED Emergency Release Button (ECR) Battery Compartment for Emergency 11 Release Battery – (PP9) 12 DVI-D Connector for External Monitor 13 14 15 16 17 RJ45 Connector (Network) Rear USB Connectors Voltage Input Label Unit Air Ex
Page |8 The xxpress unit is designed to be used with a range of sample plates and accessories – some of which are listed below: Table 3: xxpress Sample Plates and Accessories Picture Description BJS xxplate 24 well (sample volume 10-40 uL) BJS xxplate 54 well (sample volume 4-15 uL) BJS xxplate 96 well (sample volume 1-5 uL) xxsealer xxpress centrifuge Other specialist accessories may be available at request. Please contact your distributor for details. XXPRESS USER MANUAL - 1.
Page |9 4 Technical Specification Model Type Model Name Model No xxpress XP-1001-01-1 Physical Unit Properties Dimensions (W x D x H) Weight 300 mm x 520 mm x 315 mm 35 kg Mains Supply Power Cord Rating Inlet Module Type Supply Voltage Supply Frequency Range Power Consumption Fuse Ratings and Sizes Mains Supply capability Operating Environment Temperature Range Relative Humidity Range Maximum Operating Altitude Storage and Transportation Temperature Range Relative Humidity Range XXPRESS USER MANUAL
P a g e | 10 PCR Performance Thermal Ramp Rate Maximum Ramp Rate 10°C per second Adjustable from 0.1°C to 10°C per second in 0.1°C steps. Thermal Uniformity ±0.3°C well to well across the test plate Uniformity achieved after 0.
P a g e | 11 5 Safety Precautions and Limitations of Use It is essential that all users of this equipment have fully read and understood the following safety precautions and limitations of use before installing or operating the xxpress unit. IMPORTANT The protection provided by this equipment may be impaired if it is not used in a manner described in this manual. WARNING It is essential that the user of this equipment is aware of the potential hazards associated with the unit and its accessories.
P a g e | 12 Unit Installation and Operating Environment The xxpress unit is designed for indoor laboratory use only. DANGER WARNING The acceptable operating temperature range is 18ºC to 30ºC, with a relative humidity of 20% to 80% non-condensing, at a maximum altitude of 2000m above sea level. If the unit is stored in conditions outside of these ranges, it must be left to stand unpowered until it has acclimatised to within these environmental limits before being powered.
P a g e | 13 General Operating Precautions DANGER Ensure that the power is switched off at both the AC mains supply outlet and at the back of the unit before inserting or removing the mains power cord. The xxplate can reach temperatures of over 100°C and can remain hot for a time after the xxpress system is turned off.
P a g e | 14 Unit Maintenance and Serviceability There are no user or operator serviceable parts inside the unit. DANGER Do not remove the unit casework. Removal of the unit's casework will void the manufacturer’s warranty and may expose the user to a Risk of Electric Shock resulting in serious injury or death. WARNING DANGER Once installed, the externally accessible unit fuse will only blow under a fault condition.
P a g e | 15 6 Unit Installation Before installing the xxpress unit, please check that the delivery is complete and that the unit and any accessory parts are intact and free from any signs of transportation damage. Also ensure that all external and internal packaging has been removed from the unit before installation. Please retain all packaging for future transportation and storage of the unit and its accessories.
P a g e | 16 Install and test the xxpress unit using the following procedure: 1) Place the unit on the suitably selected working surface (as specified above), ensuring that the ventilation holes underneath and on the rear of the unit are not obstructed. 2) Connect the unit to the AC mains power outlet using the mains power cord supplied or as specified in Section 4. 3) Switch the mains power on at supply outlet first, and then switch the unit on using the power switch located at the rear of the unit.
P a g e | 17 7 Unit Operation WARNING Please ensure that you have read and fully understood all of the Safety Precautions and Limitations of Use listed in Section 5 before attempting to operate the xxpress unit. 7.1 User Touch Screen Display and Controls The unit’s user interface consists of a large high definition colour LCD display which has an integrated touch screen that will support multifinger gestures. (The touch screen is designed to operate when wearing standard laboratory gloves.
P a g e | 18 • The unit is now powered and so the rear fan may operate depending on the temperature of the unit and the environment. • To wake-up xxpress press the blue Power On/Off Button on the front panel once. xxpress will wake up, run through a self-test and initialise its systems. Once a screen like the one in Figure 1 is visible the xxpress is ready to use. This is the HOME screen and can always be returned to by touching the logo at the bottom left of the other screens.
P a g e | 19 Figure 2 Close the screen by touching the symbol in the top right corner of the screen. The next sections of this manual will guide the user through the regular operations that can be completed using xxpress. 7.2.2 Designing a New PCR Experiment • To create a new test method or experiment touch the DESIGN EXPERIMENT square or tile on the screen (xxpress is fitted with a touch screen, it is quite sensitive and doesn’t need high force to operate).
P a g e | 20 thermal cycling profile shown in later screens to match the experimental need). Figure 3 • In this example we have selected a 3-Step Real-Time PCR. • You will also notice running along the bottom of the screen is a navigation bar similar to that often seen on a web-site. The highlighted cyan text indicates the current stage or screen.
P a g e | 21 • The first selection is to choose between a PROBE based test or one the uses SYBR GREEN based signalling, this information enables xxpress to preselect the camera sensitivity. Touch the selection required. Figure 4a • Here we have selected SYBR GREEN see Figure 4a. • Next there are two specific selections that identify the chemistry supplier and the specific product that will be used, (this information is used to prepopulate the thermal profile with a suggested set of parameters).
P a g e | 22 • Once the chosen supplier and type of chemistry is selected (Figure 4b) then the user proceeds to the next screen by touching the arrow at the bottom right of the screen as before. Figure 4b • The next screen controls the thermal profile required for the experiment, see Figure 5. From the information already input into xxpress the system pre-populates this screen with the correct experiment structure and recommended starting values. Figure 5 XXPRESS USER MANUAL - 1.
P a g e | 23 • Looking at our example 3-Step PCR, we chose a SYBR Green based test and Kapa SYBR 2G Fast Hot Start chemistry. In Figure 5 the thermal profile reflects this with an initial hot start at 95°C for 20 seconds followed by a 3-Step cycle which is repeated 40 times. Finally there is a cooling step to bring the plate down close to ambient enabling safe removal from the unit. • Touching each part of the profile will pop-up a screen allowing the parameters to be modified, see Figure 5a.
P a g e | 24 • In the third step the temperature is increased to 72°C, the ramp rate is again 10°C per second. Once the temperature is at 60°C then this value is held for 5 seconds. At the end of this dwell there is a fluorescence measurement made using the channel 1 filter settings. • The temperatures for each step can be specified anywhere between 40°C and 99°C in 0.1°C steps.
P a g e | 25 Figure 5b • Touch before. the tick or cross to accept or reject the changes as • At this stage if a Melt Curve is to be run as part of the experiment it can be added as an additional cycle. It will then be run immediately after the PCR, see Figure 5c. BJS recommend this approach, see section 7.2.3 for how to set-up a Melt. Figure 5c • Touch the plus symbol to add the extra cycle, see Figure 5c XXPRESS USER MANUAL - 1.
P a g e | 26 Figure 5d • Touch the MELT tile to add a Melt Curve cycle to the experiment, see Figure 5d. • The Melt cycle is then added to the list of cycles and can be edited by touching. See section 7.2.3 for how to set-up a Melt Figure 5d • Once all the conditions are as required touch the forward arrow on the bottom right of the screen to advance to the Tray Selection. XXPRESS USER MANUAL - 1.
P a g e | 27 Figure 6 • Chose the desired test plate size by touching it. xxpress has a range of three standard test plates called xxplates, 24 well, 54 well, and 96 well. See Table 3 for the sample volumes they can accept. • Touch the forward arrow at the bottom right of the screen as before once the selection is correct. (xxpress will recognise the xxplate size so the correct selection is important.) Figure 7 • Figure 7 shows the chosen xxplate with its wells and the grid they are on.
P a g e | 28 • These wells can be labelled with information to help the analysis of the results. To select a group of wells to edit touch them individually or draw your finger over them. Touching them again will de-select them, or touch DESELECT ALL to clear the complete selection. Figure 7a • With the required wells highlighted touch EDIT and a screen will pop-up, see Figure 7b. Figure 7b • In the screen that pops up the type of sample that is to be put in the selected wells can be chosen.
P a g e | 29 REFERENCE GENE a sample of known response for comparison. STANDARD CURVE where that process is being completed • The display colours can also be chosen for the various charts, xxpress suggests a colour, red as shown in the example in Figure 7b. To change the suggested colour touch the coloured rectangle and further pop-up screen will appear. Touching the desired colour select it. See Figure 7c. Figure 7c • The samples can also be individually named.
P a g e | 30 • Touch the tick will accept the information and then the process can be repeated as necessary to label the rest of the plate. • You can clear the well selection or even reset the plate completely using the tiles on the right. Figure 7e • To make selection of the wells easier on the 54 well and the 96 well plates then two fingered gestures can be used to zoom into the plate area and pan around it.
P a g e | 31 • Once all the information is entered as shown in Figure 7f, touch again the forward arrow at the bottom right of the screen to move to the review screen. Figure 8 • Figure 8 shows the confirmation screen that gives the user the opportunity to review their experiment prior to running it. If anything needs editing then touching the blue navigation bar on the bottom will take the user back to the earlier screens so they can be edited or checked as required.
P a g e | 32 Figure 9 • Once the experiment is initiated xxpress performs a number of checks on the plate and initialises it systems. There will then be a short whirring sound as it clamps the plate into the unit before it starts the heat cycling. • Once the measurement cycles are running the results will appear on the chart, the chart can be zoomed by gestures as described earlier. The chart data can also be adjusted selecting the filters on the right, touch the tiles to toggle these filters on and off.
P a g e | 33 Figure 9a 7.2.3 Designing a New Melt-Curve Experiment • Creating a Melt-Curve Experiment is very similar to creating the PCR Experiment. Select MELT from the PCR menu. Figure 10 • Touch the arrow at the bottom right corner to advance to the Chemistry screen. XXPRESS USER MANUAL - 1.
P a g e | 34 Figure 11 • The Chemistry screen is simpler as just the signalling type needs to be defined to allow xxpress to set the sensitivity of the camera. • Advance to the Thermal Profile screen using the arrow as before. Figure 12 • In the Thermal Profile screen the cycle will show the range of the melt and the user can adjust that and the number of measurements taken per degree. XXPRESS USER MANUAL - 1.
P a g e | 35 Figure 12a • The example in Figure 12 above shows a Melt that will start at 40°C after a dwell of 5 seconds at 40°C. It will then rise to 95°C taking 1 measurement per degree. • The more measurements per degree greater the resolution on the melt temperatures and the longer the experiment will take. • Advance to the Tray Selection screen using the arrow as before.
P a g e | 36 Figure 14 • The plate layout can be created as shown earlier or it can be recalled from the memory. Touch the tile LOAD PLATE LAYOUT to select a previously created plate layout. • Touch the desired layout and then accept the selection by touching the tick. Figure 14a • Advance to the Confirm and Run screen using the arrow as before. XXPRESS USER MANUAL - 1.
P a g e | 37 Figure 14b • Figure 14b shows the confirmation screen that gives the user the opportunity to review their experiment prior to running it. If anything needs editing then touching the blue navigation bar on the bottom will take the user back to the earlier screens so they can be edited or checked as required. • Once the user is happy that the experiment is correctly defined then they can add the prepared test plate (see Section 7.2.4) with the test samples in.
P a g e | 38 7.2.4 Running the Experiment • Before running an experiment the sample xxplate needs to be prepared. All xxplates required to have a heat sealed film applied to them to seal the contents and prevent evaporation during the experiment. Details of how to seal the xxplates can be found in the xxsealer manual. • NEVER PUT A XXPLATE IN THE XXPRESS SYSTEM WITHOUT IT BEING HEAT SEALED. Only heat seal the xxplates with the recommended heat sealing film. • NEVER PUT A LABEL ON THE BOTTOM OF THE XXPLATE.
P a g e | 39 • Place the xxplate onto the slides ensuring that it correctly located so that the slides engage into all three holes on either side of the xxplate, see Figure 15. • When correctly located close the door by pushing on the front of it until it latches shut. • Once the experiment has been started by touching the forward arrow on the RUN screen then the xxpress system will run a number of checks on the plate. These are listed in an information window that pops up in the middle of the display.
P a g e | 40 • During the operation it is normal for the unit to issue a short hum from the cooling compressors every cycle as they are used to cool the plate • Once the experiment is complete the clamps will release and an information window will pop up to confirm this. Figure 16a 7.2.5 Experiment Results and Analysis • At the end of an experiment the results are displayed as shown in Figure 17. Depending on the type of experiment different parameters are shown.
P a g e | 41 • There is a manual adjustment that can be used to adjust the Ct threshold value by eye. Touch the red circle and slide it up and down to adjust the threshold, the value is shown in the menu bar on the right of the screen. Figure 17 • When looking at a Melt there is the option of looking at the differential of the curve which helps identify the Tm more easily. Figure 17a shows an example of a Melt in it standard form. Figure 17a XXPRESS USER MANUAL - 1.
P a g e | 42 • Touching the DIFFERENTIATE tile on the right will give a display like the one shown in Figure 17b. Figure 17b • Results can be export in a CSV format so they can be analysed in other ways as required. Select the blue EXPORT TO CSV tile to do this. Note, if the smoothing filter or background filter is active when the data is exported the values will include this filtering.
P a g e | 43 8 Maintenance and Servicing Although the xxpress unit does not require any scheduled maintenance or servicing, the operator should regularly clean and inspect the unit for any detects, as described in Section 8.2 below. DANGER WARNING Please observe and comply with all of the Unit Maintenance and Serviceability precautions listed in Section 8. Never removed the unit casework. There are no user or operator serviceable parts inside the unit.
P a g e | 44 Figure 18 1. Remove the AC plug from the power inlet module 2. Use a small flat bladed screwdriver to carefully pull out the fuse holder 3. Fit the correct fuse in the far position only 4. Push the holder back into the inlet mode 8.2 Routine Cleaning and Inspection The unit casework should be cleaned and inspected at regulator internals, and whenever contamination or spillage occurs, as follows: 1.
P a g e | 45 8.3 Decontamination Procedure The unit and accessories should be decontaminated using the following procedure before being stored or transported. Decontamination Procedure Thoroughly clean all outside surfaces of the product (including any accessories, power cords, manuals, packaging, etc) with a damp cloth soaked with suitable disinfectant solution (such as Virkon). 8.
P a g e | 46 Certificate of Decontamination We respect the health and safety of our clients and employees, and request that any products or accessories being returned are decontaminated in accordance with the procedure below. 1. Decontamination Procedure Thoroughly clean all outside surfaces of the product (including any accessories, power cords, manuals, packaging, etc) with a damp cloth soaked with suitable disinfectant solution (such as Virkon). Allow to dry fully before packing. 2.
P a g e | 47 9 Warranty and Returns BJS Biotechnologies Ltd. warrants the xxpress product, when purchased new and installed and operated in accordance with the instructions of this manual, to be free from defects in materials and workmanship, and will repair or replace, at their discretion, any unit or accessory which exhibits such defects. In no event will BJS Biotechnologies be liable for any indirect, incidental or consequential damages resulting from any defect or warranty claim.
P a g e | 48 10 Product Disposal At end-of-life, this product must be disposed of in accordance with your local authority regulations for the disposal of potentially hazardous waste and electronic equipment. The unit and its accessories should be decontaminated using the procedure detailed in Section 8.3 before disposal or shipping. Do not dispose of this product into unsorted municipal waste or public landfill.
