The World’s F
astest Real-Time PCR
Thermal Cycler
User Manual
This Manual Copyright © 2018 BJS Biotechnologies Ltd.
Страница 1: ...The World s Fastest Real Time PCR Thermal Cycler User Manual This Manual Copyright 2018 BJS Biotechnologies Ltd...
Страница 2: ...d BJS Bio The information contained in this manual is subject to change without notice BJS Bio assumes no responsibility for any errors that may appear in this or related documentation Document XP 500...
Страница 3: ...urning on xxpress NGx 18 7 2 2 Designing a New PCR Experiment 20 7 2 3 Designing a New Melt Curve Experiment 37 7 2 4 Running the Experiment 41 7 2 5 Experiment Results and Analysis 43 7 2 6 Repeating...
Страница 4: ...Chain Reaction PPE Personal Protective Equipment Real Time PCR Polymerase Chain Reaction where the result is delivered throughout the PCR process by the capturing of fluorescence emitted by the reacti...
Страница 5: ...Indicates a Risk of Explosion which could if not avoided result in serve injury or death WARNING Indicates a hazardous situation which could if not avoided result in serve injury or death or severely...
Страница 6: ...evel of fluorescence emitted by the samples xxpress NGx has five fluorescence measurement channels that enable fluorophores with a wide range of excitation and collection frequencies to be detected To...
Страница 7: ...Touch Screen 6 Front USB Connector 7 Rear Cover 8 Air Intake Vents and Filters under unit 9 Power on LED 10 Emergency Release Button ECR 11 Battery Compartment for Emergency Release Battery PP9 12 DV...
Страница 8: ...e listed below Table 3 xxpress NGx Sample Plates and Accessories Picture Description 24 well xxplate SBS sample volume 5 40 L 54 well xxplate SBS sample volume 5 15 L 96 well xxplate SBS sample volume...
Страница 9: ...n Only use an appropriately rated and approved supply cordset in accordance with the regulations of the country in which the equipment is put into service IEC C14 DPST Single Fuse 100V 120V AC XP 1002...
Страница 10: ...om 0 1 C to 10 C per second in 0 1 C steps 0 3 C well to well across the test plate Uniformity achieved after 0 5 seconds 5 Channel Fluorescence Detection LED illumination CCD image capture Detection...
Страница 11: ...d be familiar with the safety precautions and warnings given in these instructions before attempting to operate the unit Improper use of this unit or its accessories may impair their functionality and...
Страница 12: ...urely inserted into the rear of the unit and any excess power cord does not pose a potential trip or pull hazard Always ensure that the mains plug is easily accessible to allow disconnection from the...
Страница 13: ...the xxpress NGx system is turned off If conditions are set such that the experiment is finished with the plate at a high temperature it will exit the unit with the potential to burn the operator and...
Страница 14: ...ineer See Section 8 1 for details DANGER Always switch off the unit and disconnect the power cord before performing any cleaning or decontamination procedure If liquid is spilt into or over the unit s...
Страница 15: ...orking surface At least 10cm clearance around the unit to adjacent objects and walls Earthed AC mains power connection see Section 4 WARNING Please also observe and abide by the Unit Installation and...
Страница 16: ...s shown below Close the battery compartment 3 Connect the unit to the AC mains power outlet using an appropriately rated and approved cordset as detailed in Section 4 4 Switch the mains power on at th...
Страница 17: ...P a g e 17 XXPRESS NGX USER MANUAL XP 5001 01 5 6 Start up is complete when the LOGON screen is shown see below...
Страница 18: ...been designed to match experimental procedures used by Biotechnologists making it intuitive and easy to use The xxpress NGx software and user instructions will be regularly updated You can find detai...
Страница 19: ...the logo in the bottom left corner of the screen From the HOME screen it is possible to create new experiments run previously designed experiments and analyse the results of previously run experiments...
Страница 20: ...t method or experiment touch the DESIGN EXPERIMENT tile on the screen the xxpress NGx is fitted with a touch screen which is sensitive and doesn t need significant force to operate The next screen ask...
Страница 21: ...o match the experimental need Figure 3 In this example we have selected a 3 Step Real Time PCR You will notice that there is a navigation bar running along the bottom of the screen The highlighted cya...
Страница 22: ...the camera sensitivity Figure 4a In Figure 4a SYBR GREEN has been selected This brings up a list of some kit manufacturers that have been tested by BJS Bio Touch to select which kit manufacturer is be...
Страница 23: ...d chose a similar product as a starting point and edit the thermal parameters at a later stage With subsequent software releases BJS Bio will add new chemistry options Between releases application not...
Страница 24: ...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 fol...
Страница 25: ...s raised to 95 C using a ramp rate of 10 C per second This temperature is then held for 1 second In the second step the temperature is reduced to 60 C using a ramp rate of 8 C per second Once the temp...
Страница 26: ...al segment sees the temperature being reduced to 50 C and held for 10 seconds This is done to cool the plate enough to touch when removed from the xxpress NGx Based on the chemistry selection xxpress...
Страница 27: ...espectively 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 a...
Страница 28: ...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 0 for how to set up a Melt Figure 5d Once all the parameters have be...
Страница 29: ...it The xxpress NGx has three xxplate sizes 24 well 54 well and 96 well See Table 3 for the reaction volumes they can accept Touch the forward arrow at the bottom right of the once the selection is co...
Страница 30: ...the wells can be labelled with information to help with the results analysis To edit a group of wells touch to select them see Figure 7a Touching them again will de select them alternatively you can t...
Страница 31: ...the display colours for the amplification and melt graphs The xxpress NGx will suggest a colour but this can be changed Touch the coloured rectangle and a pop up window will appear as shown in Figure...
Страница 32: ...et the plate completely using the tiles on the right Figure 7e To make the selection of smaller wells easier i e when using a 54 well xxplate or 96 well xxplate use two fingers to zoom into the plate...
Страница 33: ...n is entered as shown in Figure 7f touch the forward arrow at the bottom right of the screen to move to the review screen Figure 8 Figure 8 shows the review screen This gives the user the opportunity...
Страница 34: ...8a NEVER PUT A XXPLATE IN THE XXPRESS NGx SYSTEM WITHOUT IT BEING HEAT SEALED Only heat seal the xxplates SBS with the recommended heat sealing film NEVER PUT A LABEL ON THE BOTTOM OF THE XXPLATE Keep...
Страница 35: ...he graph data can be adjusted using the filters on the right Touch the tiles to toggle the filters on and off BASELINE SUBTRACT uses the fluorescence readings of the first 5 cycles to set the initial...
Страница 36: ...P a g e 36 XXPRESS NGX USER MANUAL XP 5001 01 5 automatically and can now be analysed and or exported see Section 7 2 5 Figure 9a...
Страница 37: ...CR Experiment Select MELT from the PCR menu see Figure 10 Figure 10 Touch the arrow at the bottom right corner to advance to the Chemistry screen Figure 11 The Chemistry screen is simpler Only the sig...
Страница 38: ...mber of measurements taken per degree Figure 12a The example in Figure 12a above shows a Melt that will hold at 40 C for 5 seconds before raising the temperature from 40 C to 95 C taking 1 measurement...
Страница 39: ...13 Figure 13 Advance to the Plate Layout screen using the arrow Figure 14 This is an optional page A plate layout can be created as shown earlier or alternatively it can be recalled from the memory To...
Страница 40: ...see Figure 14a Figure 14a Advance to the Confirm and Run screen using the arrow Figure 14b Figure 14b shows the confirmation screen that gives the user the opportunity to review their experiment prio...
Страница 41: ...the Experiment Before running an experiment the sample xxplate needs to be prepared All xxplates are required to have a heat sealed film applied to them This seals the contents and prevents evaporati...
Страница 42: ...been started by touching the forward arrow on the RUN screen the xxpress NGx will run a number of checks on the plate These are listed in an information pop up window on the display These include the...
Страница 43: ...displayed as shown in Figure 17 Depending on the type of experiment different parameters are shown For a PCR run the Cq value is calculated whereas for a MELT the Tm value is calculated The results gr...
Страница 44: ...nd shown in for each well in the ANALYSIS tab see Figure 17a The threshold for the Cq measurements is also calculated and is shown in a blue box on the PCR tab This can be manually adjusted by sliding...
Страница 45: ...melt section you have the option to differentiate the graph This helps identify the Tm more easily Figure 17b shows an example of a Melt in it standard form Figure 17b Touching the DIFFERENTIATE tile...
Страница 46: ...format ready for further analysis Select the blue EXPORT 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 S...
Страница 47: ...uorescence channel the data can be viewed using the coloured channel tiles at the bottom of the screen see Figure 17e Where there may be cross talk between adjacent fluorescence channels touching the...
Страница 48: ...e The experiment will open at the Confirm and Run screen see Figure 18a Figure 18a Touching the menu along the bottom of the screen enables the selection of any of the experiment set up screens These...
Страница 49: ...y run Figure 18b All the results from the experiments that have been completed are stored on the xxpress NGx These can be reviewed at any time To review the results of a completed experiment touch the...
Страница 50: ...replaced by a suitably qualified technician DANGER The unit fuse will only blow as a result of an internal unit fault or if the voltage selector switch has been incorrectly set see Section 5 This fuse...
Страница 51: ...efore cleaning always inspect the unit casework heater plate and moving parts for any signs of wear damage cracks or other defects 3 Clean the casework using a damp cloth soaked with a disinfectant so...
Страница 52: ...etc with a damp cloth soaked with suitable disinfectant solution such as Virkon 8 4 Transportation and Storage The xxpress NGx unit and its accessories should be thoroughly decontaminated using the p...
Страница 53: ...ing 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 Decontamination Declaration Compan...
Страница 54: ...eral not supplied specified or sanctioned by BJS Biotechnologies will invalidate this warranty This warranty is provided to the original purchaser of the product for one year from the date of purchase...
Страница 55: ...tially 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 pro...
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Страница 57: ...BJS Biotechnologies Ltd 65 Bideford Avenue Perivale Greenford Middlesex UB6 7PP United Kingdom www xxpressPCR com...
