Thermo Hybaid PX2 User Instruction Manual Download Page 53

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CHAPTER 7

Px2

THERMAL CYCLER

Troubleshooting

7.1 Programs Using ACTIVE Tube Control

Check the Volume in the Tube Thermistor

Check the volume of mineral oil in the control tube (a vial of suitable mineral oil is provided

(alternatively Sigma molecular biology-grade mineral oil, M5904 or equivalent can be used).

Do not

use aqueous solutions in the control tube as this can reduce the life expectancy of the

thermistor.

Too much oil in the control tube will result in overshoots in the sample tube temperatures,

whilst too little oil there will result in a time lag in the samples achieving temperature. The volume of oil

in the control tube should match that in the reaction tubes (including any oil overlay). Repeated

overshoots at the denaturation temperature during temperature cycling will reduce the activity of the

thermostable enzyme resulting in poor yields of product. The correct position for the control tube is

position A2.

Check Location of Thermistor within the Control Tube

Always ensure the thermistor probe is located centrally in the control tube and immersed in the liquid.

If pushed against the side it will be measuring the temperature of the microcentrifuge tube and not the

sample temperature.

Check the Fit of the Tube in the Block

The Px2 block accommodates most types of reaction tubes. The tube thermistor supplied with the

Px2 is mounted on an OmniTube, which should be changed periodically, as the fit of the tube will

deteriorate with time. Reaction tubes should be distributed evenly in the block. If you are using a

reaction tube that has not been recommended, check its fit in the Px2 block before use. You should

also remount the thermistor probe in this tube to ensure that your samples and the control tube are

matched.

Check through the Program

Before running a program, use the VIEW function to check that the correct combination of

temperature and program number have been entered.

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Summary of Contents for PX2

Page 1: ...l service in house repair center WE BUY USED EQUIPMENT Sell your excess underutilized and idle used equipment We also offer credit for buy backs and trade ins www artisantg com WeBuyEquipment REMOTE INSPECTION Remotely inspect equipment before purchasing with our interactive website at www instraview com LOOKING FOR MORE INFORMATION Visit us on the web at www artisantg com for more information on ...

Page 2: ...ermo Hybaid Action Court Ashford Road Ashford Middlesex TW15 1XB Telephone 44 0 1784 425 000 Facsimile 44 0 1784 248 085 Internet www thermohybaid com STOCK CODE HB PX2 MAN FS31999 Artisan Technology Group Quality Instrumentation Guaranteed 888 88 SOURCE www artisantg com ...

Page 3: ... Thermo Hybaid Issue 1 0 January 2002 ii Artisan Technology Group Quality Instrumentation Guaranteed 888 88 SOURCE www artisantg com ...

Page 4: ...trument is also an Authorized Thermal Cycler for use with applications licenses available from Applied Biosystems Its use with Authorized Reagents also provides a limited PCR license in accordance with the label rights accompanying such reagents Purchase of this product does not itself convey to the purchaser a complete license or right to perform the PCR process Further information on purchasing ...

Page 5: ... Issue 1 0 January 2002 iv Key Pad Chassis Main Body Stabilising Feet Heated Lid Height Selector Heated Lid Large LCD Screen Artisan Technology Group Quality Instrumentation Guaranteed 888 88 SOURCE www artisantg com ...

Page 6: ...ybaid s option Thermo Hybaid is not liable for any consequential damages The tube thermistor assembly supplied with your Px2 unit is guaranteed for 90 days Thermo Hybaid reserves the right to alter the specification of the Px2 without prior notice This will enable us to implement developments as soon as they arise The Thermo Hybaid Px2 is for research use only Read the Instruction Manual carefully...

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Page 8: ... 2 Heating and Cooling 2 Programming and Operation 3 Pre set Programs 3 Advanced Edit Temperature Ramping 3 Advanced Edit Time Temperature Increment 3 Advanced Edit Gradient optional 4 Calibration of your Px2 4 1 3 The Heated Lid 4 Operating the Lid 4 1 4 Input Output Sockets 6 1 5 Support Services 6 CHAPTER 2 SAFETY PRECAUTIONS 7 2 1 General Safety Precautions 7 2 2 Symbols and Conventions 8 CHAP...

Page 9: ...unning a Program 16 4 7 Manual Operation 16 4 8 Programming Function 17 4 9 Loading Samples and Heated Lid Operation 18 CHAPTER 5 PROGRAMMING THE Px2 19 5 1 Introduction 19 5 2 Directories 19 Creating a New Directory 19 Editing a Directory Name 20 Erasing an Existing Directory 20 5 3 Programs 21 Creating a New Program 21 Editing an Existing Program 22 Viewing an Existing Program 24 Copying Program...

Page 10: ... Options 33 Active Tube Control ACTIVE TUBE 33 Extension Lead for Tube Thermistor 34 Simulated Tube Control SIM TUBE 35 Simulated Plate Control SIM PLATE 35 Simulated Slide Control SIM SLIDE 36 Block Control 37 6 5 Heated Lid Preheat 37 Heated Lid Automatic Start 37 Heated Lid Manual Start 37 No Heated Lid 38 6 6 Hold and Pause Function 38 6 7 Aborting Programs 38 6 8 Program Completion 39 CHAPTER...

Page 11: ...nealing 44 Extension 45 CHAPTER 8 MAINTENANCE 47 8 1 General Cleaning 47 8 2 Decontamination 47 8 3 Protection to the User 48 8 4 Protection of the Instrument 48 Fuses 48 8 5 Tube Thermistor Care 48 CHAPTER 9 TECHNICAL SPECIFICATIONS ORDERING INFORMATION 51 APPENDIX I RECOMMENDED CONSUMABLES 57 APPENDIX II SEALING SYSTEMS THE OMNISEAL TD MAT TD TAPE 59 APPENDIX III IN SITU BLOCK APPLICATION NOTES ...

Page 12: ... can be changed according to the needs of the sample format There are six types of block module The 0 2ml Standard and Gradient Block Module This block can hold any of the following 1 x OmniUtra 96 well skirted polycarbonate plate 1 x OmniFast 96 well skirted polypropylene plate 1 x OmniTube 96 single piece polypropylene plate 96 x 0 2ml OmniStrip tubes 96 x 0 2ml individual OmniTubes The 0 5ml St...

Page 13: ...r 384 Well OmniPlates or racked OmniTubes The temperature control algorithm is similar to active tube control but is based on calculated values for the sample temperature rather than values fed back by the tube thermistor 3 Simulated Slide Control For the in situ module Simulated Slide Control should be selected This operates on a similar principle to Simulated Tube Control but in this case the al...

Page 14: ...ling techniques and can be used to create customised protocols by editing see Appendix V Advanced Edit Temperature Ramping The temperature ramping Ramp Rate enables the rate of change of sample temperature C second to be artificially slowed down Under normal circumstances the temperature cycling times are very rapid which minimises non specific reactions The rate of sample temperature change durin...

Page 15: ...al protocol is largely unchanged when switching to an oil free format The heated lid operates by positioning a heated plate in contact with the top of the reaction tubes or wells This heated plate then heats the air temperature at the top of each reaction mixture to a temperature that is permanently higher than the sample temperature This elevated air temperature relative to the sample temperature...

Page 16: ...er it is a plate or not Please note that the tube thermistor must be removed before selecting the plate mode Figure 1 1 Open Lid Icon Tube Icon Plate Icon The temperature of the heating plate is set at 120 C max surface temperature with power control effected by the control chassis The heated plate inside the lid is switched on and off from within the program The lid operates once a program is act...

Page 17: ...ack of the unit corresponds to RS485 1 5 Support Services The Px2 has been designed for reliability and for ease of maintenance Thermo Hybaid continues to offer full service and technical support for all its products Thermo Hybaid Head Office Tel 44 0 1784 425 000 Fax 44 0 1784 248 085 Email globalsupport thermohybaid com Alternatively appropriate contact details for your local Thermo Hybaid subsi...

Page 18: ... avoid the risk of personal injury and to ensure that the safety features of this unit are maintained servicing should only be carried out by authorised service personnel 8 The tops of the reaction vessels and the surfaces of the heated lid assembly in particular the inner surfaces can become very hot during normal operation Touching the surfaces can cause burns Do not touch the heated plate witho...

Page 19: ...Proceed with caution to avoid burn injury ATTENTION Surface chaude SAFETY NOTE This symbol indicates high voltage Risk of electric shock AVERTISSEMENT Risqué de choc éléctrique In addition the following conventions are adopted in the manual in respect of indicating safety hazard SAFETY NOTE indicates a potentially hazardous situation which could result in death or serious injury WARNING indicates ...

Page 20: ... to stabilise at normal ambient temperature before powering up the unit 3 2 Packing List 1 Px2 Control Chassis 2 Px2 Block Module optional 3 User Instruction Manual 4 Mains Lead 5 Control Thermistor Probe 0 2ml and 0 5ml gradient and standard block modules 6 Control Thermistor Probe Extension Lead 0 2ml and 0 5ml gradient and standard block modules 7 Specific Consumable Pack 8 RS232 Lead NB The In...

Page 21: ...ndoor use at an ambient temperature of 4 34 C in conditions of up to 80 humidity These specifications have been calculated for operations at between 0 and 2000m altitudes Block Installation and Removal 1 Remove the Px2 Control Chassis and Block Module from the packaging and place on the bench do not connect to the mains supply immediately 2 Open the lid by turning the knob anticlockwise to the ope...

Page 22: ...hat the probe is immersed in the oil and not touching the wall of the reaction vessel Figure 3 2 Well Numbering Showing Tube Thermistor Positioning in A2 Use in Cold Rooms The Px2 may be used in a cold room down to 4 C However when removed to room temperature the instrument should be allowed to equilibrate for at least two hours before being switched on If equilibration is not done condensation ma...

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Page 24: ...any time by pressing MENU The Main Menu displays the date time and the status of the block active inactive or active resumed All aspects of the programming are accessed through the Main Menu Table 4 1 Summary of Main Menu Functions RUN Select RUN to run an existing program MAN Select MAN to run a manual static incubation PROG Select PROG to edit copy view or erase a program OPTS Select SET UP to a...

Page 25: ...ed to toggle between options 4 4 Set Up Functions Selecting OPTS at the Main Menu by pressing the relevant soft key gives access to the SETUP menu Within the SETUP screen information about the type of block fitted the current software version installed the time and the date are displayed A flashing cursor beneath CLOCK now indicates that this application will be selected when ENTER is pressed The ...

Page 26: ...rded 4 5 Gradient Calculation Once you have run your samples on an agarose gel the annealing temperature that corresponds to your optimum gel result can be determined using the Gradient Calculator function To access the Gradient Calculator select OPTS from the MAIN MENU Using the left and right navigation keys move the cursor to CALC and press ENTER The block type you are using will automatically ...

Page 27: ...VIOUS and NEXT keys to change to the appropriate method A BACK button can be used to return to the previous screen if necessary Pressing ENTER will now start the program Prior to starting the program pressing the MENU key at any stage will return to the Main Menu More details can be found in Chapter 6 4 7 Manual Operation This option is used for static incubations when thermal cycling is not requi...

Page 28: ...otocols of varying degrees of complexity can easily be created using Px2 As with all operations of the Px2 programming is accessed from the Main Menu Access the Program Menu by selecting PROG from the main menu If at any time you wish to stop programming press MENU to return to the Main Menu Four options are available for programming The LEFT or RIGHT arrow keys are used to move the cursor to the ...

Page 29: ...ock The heated lid is simple to operate In one action the lid is closed and the heated plate lowered to its correct height and pressure setting The pressure exerted by the lid can be adjusted for tubes or plates by rotating the knob so that the relevant icon is aligned to the front of the unit see Figure 4 1 It is defaulted to the tube pressure for the plate pressure turn the knob clockwise until ...

Page 30: ...simply i Enter or create a new directory ii Name a program iii Enter protocol parameters iv Save the program 5 2 Directories There are five directories A E which can be individually named The sixth directory F is reserved for pre set programs Directories can also be erased and renamed as required Once assigned to a directory a program can be copied to another program space in either the same or a ...

Page 31: ... The cursor should be flashing below EDIT select this option by pressing ENTER 3 Use the RIGHT softkey to move the cursor to NEW and select by pressing ENTER 4 Use the RIGHT softkey to move the cursor to NEW DIR and select by pressing ENTER 5 Use the UP and DOWN cursor to select the directory you wish to rename and press ENTER 6 Use the alphanumeric keypad to enter the new directory name Use the P...

Page 32: ...speeds setting a gradient of annealing temperatures changing time temperature with successive cycles can be accessed by pressing the ADVANCED softkey at the relevant step see section 5 4 for further details Creating a New Program 1 From the Main Menu select PROG using the appropriate softkey 2 Move cursor to EDIT then press ENTER to select 3 Move the cursor to NEW then press ENTER to select 4 Move...

Page 33: ...e number of times you would like to cycle through the STAGE using the alphanumeric keys A hold step can also be programmed after each stage so once the cycles have been completed the Px2 will wait at the specified temperature until manually advanced This is particularly useful for hot start protocols If a hold step is not required press ENTER to reach the next stage 10 Continue adding steps to the...

Page 34: ...3 Select EXISTING from the Program Editing Menu by pressing ENTER 4 Use the UP and DOWN keys to move the cursor to the appropriate source directory and press ENTER to select The BACK button may be used to return to the previous screen if necessary 5 The keypad numbers or PREVIOUS and NEXT softkeys can be used to select the number of the program to be edited Press ENTER to select the program 6 Use ...

Page 35: ... select the appropriate directory and then press ENTER 3 Use the PREVIOUS and NEXT softkeys to select the program and press ENTER 4 Use the NEXT softkey to scroll through the program information Press QUIT or MENU at any time to return to the Main Menu Copying Programs Copying programs is necessary in for example modifying the pre set programs described in Appendix V 1 Select PROG from the Main Me...

Page 36: ...evant program by moving the cursor with the UP and DOWN softkeys and press ENTER 4 Use the PREVIOUS and NEXT keys to choose the relevant program and press ERASE 5 A final confirmation screen allows you either KEEP or ERASE the program 5 4 Advanced Edit Features The Advanced Edit features allow the creation of more complex cycling protocols in order to enhance experimental data It is possible to pr...

Page 37: ...each program stage Time increments can be used for example in high cycle number reactions to allow longer for enzyme action with successive cycles Below is an example of a protocol where the extension step is fixed for the first 15 cycles of a 25 cycle program and increased by 5 seconds cycle for the next 10 cycles to compensate for loss of enzyme activity Stage 1 Enter parameters to create 95o C ...

Page 38: ...ting screen ADV now appears on the screen indicating an Advanced Edit function has been programmed Temperature Advanced Edit Worked Example Temperature decrements can be used for example in touchdown cycling reactions where the annealing temperature is decreased with successive cycles This example shows a protocol where the annealing step is fixed for the first 5 cycles of a 25 cycle program and d...

Page 39: ... the maximum practical setting being 3 00o C sec with current technologies The data entered refers to the ramp rate to the CURRENT step from the PREVIOUS temperature set point For example cycle sequencing of some targets requires the use of degenerate primers and a ramp rate set at 1o C second between the annealing temperature and extension temperature 5 5 Gradient Feature This feature is only ava...

Page 40: ...n these limits E g SET TEMP 60o C and GRAD 15o C The SET TEMP will be changed to 55 C If a temperature is entered that exceeds the 15o C gradient range then a error message will appear beside the GRAD parameter A gradient range from 1 15o C will need to be re entered Gradient Calculator Function Once you have run your samples on an agarose gel the annealing temperature that corresponds to your opt...

Page 41: ...aid Issue 1 0 January 2002 30 The Gradient Calculator may be accessed at any time whether the block is active or inactive Artisan Technology Group Quality Instrumentation Guaranteed 888 88 SOURCE www artisantg com ...

Page 42: ... Thermo Hybaid Issue 1 0 January 2002 31 Artisan Technology Group Quality Instrumentation Guaranteed 888 88 SOURCE www artisantg com ...

Page 43: ... control for an in situ interchangeable block module simulated slide for an 02 05 module then a warning message will appear and the program will be run under the correct mode of control 6 2 Running a Program on the Px2 Short Instructions Temperature control and heated lid functions are referred to in more detail in section 6 4 1 Select RUN from the MAIN MENU 2 Use the UP and Down softkeys to selec...

Page 44: ...een using the LEFT softkey TEXT GRAPH These screens can be accessed from the Main Menu by pressing the RUN INFO softkey Text Screen The current block performance screen displays the following Program number and name Status of the program The temperature control mode being used Target programmed temperature for a given step The block and tube temperatures Time remaining at current setpoint The curr...

Page 45: ...atures recorded during the run Power interrupt information if a power failure occurred Error Screens A number of error screens may be displayed if a fault has been detected in the unit Contact the Thermo Hybaid Service Department or your local supplier for advice before attempting to use the instrument further 6 4 Temperature Control Options Crucial to the accurate operation of a thermal cycler is...

Page 46: ...the programmed time With block control either on the Px2 or a thermal cycler without tube control there will be a lag between the block reaching target temperature and the sample reaching target temperature Thus when transferring protocols from a block control machine the incubation times may be reduced by up to 50 and in some cases the temperatures adjusted slightly For Active Tube Control reacti...

Page 47: ...ube thermistor to be connected but sample loading and volume details must be entered when prompted by the run screen For all tube reactions in a 0 2ml or a 0 5ml thin walled tube the volume factor is the total reaction volume in µl in one well including any oil overlay Simulated Plate Control SIM PLATE This mode of control operates on a similar principle to simulated tube control see Figure 6 1 Ho...

Page 48: ...bes attached to the surface of slides Optimum results in terms of overshoot and uniformity across four standard glass microscope slides 0 8mm 1 0mm thickness with the humidity chamber in place have been obtained using a calibration factor of 25 the default value Using a higher calibration accelerates the approach to target temperature but results in some degree of overshoot Conversely a lower cali...

Page 49: ...generation of spurious products After the program is selected the Lid Heating indicator light will switch on denoting preheating of the lid When the lid reaches operating temperature the Ready Working indicator light will change to green and the temperature cycling will start automatically In this mode samples should be loaded and the lid closed before starting the preheat Heated Lid Manual Start ...

Page 50: ...of the protocol to perform the Hot Start procedure After the enzyme has been added pressing CONTINUE will advance the program into the cycling part of the protocol A final low temperature 4 10o C hold for the end of overnight runs Although unnecessary for the vast majority of protocols some scientists prefer to have this step included A final 72o C incubation to ensure completion of the final exte...

Page 51: ...r 30 minutes to drive the heated lid plate temperature to ambient If a final HOLD temperature has been specified the heated lid if selected will remain on For most thermal cycling applications a final HOLD at elevated temperature is not necessary Extended high temperature hold steps can lead to evaporation and condensation problems particularly with polycarbonate plates Artisan Technology Group Qu...

Page 52: ... Thermo Hybaid Issue 1 0 January 2002 41 Artisan Technology Group Quality Instrumentation Guaranteed 888 88 SOURCE www artisantg com ...

Page 53: ...rrect position for the control tube is position A2 Check Location of Thermistor within the Control Tube Always ensure the thermistor probe is located centrally in the control tube and immersed in the liquid If pushed against the side it will be measuring the temperature of the microcentrifuge tube and not the sample temperature Check the Fit of the Tube in the Block The Px2 block accommodates most...

Page 54: ...f the OmniPlate 96 in the 0 5ml block Other 96 well plates for example the Costar ThermowellH may be used with lower multiples of volume but these need to be individually determined 384 well blocks For 384 well blocks the individual well volume should be used for the volume factor Simulated Slide For 0 8mm 1mm thick glass slides using simulated slide control the calibration factor is 25 default va...

Page 55: ... the instrument is set to block control This enables the tube thermistor to be used remotely 2 Connect the thermistor lead extension to the thermistor control tube and plug it into the Px2 3 Place the thermistor tube probe into the corresponding thermistor probe socket of the Px2 and locate the tube in a well of the block control machine running the required protocol 4 After a short equilibration ...

Page 56: ...sation of the denaturation step is the most critical factor when transferring a protocol from a block control machine to tube control Annealing The annealing temperature depends on the size and nucleotide composition of the oligonucleotides used In general it varies between 50 C and 70 C and as a rough guide should be 5 C below the Tm This may be calculated approximately using the following formul...

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Page 58: ...tamination When 35S labelled nucleotides are thermally cycled they break down into lower molecular weight forms which are highly volatile and can leach through the walls of tubes and microtitre plates thus contaminating the block and possibly the heater plate of the heated lid We therefore do not recommend the use of 35S labels as replacing a dangerously contaminated block is expensive If 35S labe...

Page 59: ...nts 8 3 Protection to the User The Px2 has been designed with operation safety in mind In the rare event of an instrument failure three levels of protection are built in to ensure the unit fails safe First the software sets normal operating ranges for the block and lid Should this fail electrical circuitry is in place to ensure that safe temperatures are not exceeded In the unlikely event of this ...

Page 60: ...be if it is showing signs of thermal degradation 4 Do use the same type of tube for the thermistor tube as for the biological samples 5 Do make sure the sensor part of the thermistor probe is centrally located at the base of the tube and is fully covered by mineral oil 6 Don t use aqueous solutions in the thermistor tube 7 Don t remove the control tube from the cap assembly by pulling the wire gen...

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Page 62: ...rol Simulated plate control Block control Performance Block Temperature Range 4 C 99 C 4 C 99 C Block Heating Rate Up to 3 C Sec Up to 3 C Sec Block Cooling Rate Up to 2 C Sec Up to 2 C Sec Precision Control 0 1 C 0 1 C Block Uniformity 0 4 C within 15 secs 0 4 C within 15 secs Heated Lid Temperature Range 95 C 120 C 95 C 120 C Display Resolution 0 1 C 0 1 C Ingress Protection Rating 20 20 Standar...

Page 63: ... Block Temperature Range 4 C 99 C 4 C 99 C Block Heating Rate Up to 3 C Sec Up to 3 C Sec Block Cooling Rate Up to 2 C Sec Up to 2 C Sec Precision Control 0 1 C 0 1 C Block Uniformity 0 4 C within 15 secs 0 4 C within 15 secs Heated Lid Temperature Range 95 C 120 C 95 C 120 C Display Resolution 0 1 C 0 1 C Ingress Protection Rating 20 20 Standard Accessories Humidity Chamber Interchangeable Block ...

Page 64: ...ock control Performance Block Temperature Range 4 C 99 C 4 C 99 C Block Heating Rate Up to 3 C Sec Up to 3 C Sec Block Cooling Rate Up to 2 C Sec Up to 2 C Sec Precision Control 0 1 C 0 1 C Block Uniformity 0 4 C within 15 secs 0 4 C within 15 secs Heated Lid Temperature Range 95 C 120 C 95 C 120 C Display Resolution 0 1 C 0 1 C Ingress Protection Rating 20 20 Standard Accessories Tube Thermistor ...

Page 65: ...9hr 59 mins 59 secs Time increment decrement Yes Temp increment decrement Yes Temperature ramping Yes Pause facility Yes Autostart facility Yes Run end time calculations Yes File protection Yes Output RS232 and RS485 Alphanumeric programming Yes Gradient software Yes Power 550W Dimensions WxDxH 240mm x 390mm x 280mm Weight 8 6kg Artisan Technology Group Quality Instrumentation Guaranteed 888 88 SO...

Page 66: ...ments 550W at 115 230V a c 10 and 50 60Hz Satisfies the requirements of BS EN 61010 1 1993 NB When the Px2 thermal cycler is removed from a cold room it should be left to equilibrate for 2 3 hours to avoid condensation Artisan Technology Group Quality Instrumentation Guaranteed 888 88 SOURCE www artisantg com ...

Page 67: ... Thermo Hybaid Issue 1 0 January 2002 56 Artisan Technology Group Quality Instrumentation Guaranteed 888 88 SOURCE www artisantg com ...

Page 68: ...TAPE100 Compression Pad 1 Foam pressure pad for use with tape sealing system HB TD SFOAM OmniSeal TD Mat 5 Reusable 96 well sealing mat HB TD MTSRS5 Plasticware Consumables Pack Size Description Catalogue Number Plates OmniUltra Non Irradiated 25 Polycarbonate 96 well plate robot compatible HB TC 25D OmniUltra Gamma Irradiated 25 Polycarbonate 96 well plate robot compatible HB TC 25N OmniUltra Lid...

Page 69: ...662N OmniStrip 0 3ml 2000 0 3ml strips of 8 tubes caps HB TC 4043N Racked OmniTubes 25 For 0 3ml 96 well plates caps not included HB TC 4073N OmniTube Caps 2400 For all 0 2ml and 0 3ml OmniTube products HB TC 6022N We recommend that domed cap tubes are used when the temperature control is set to Active Tube and flat cap tubes are used under Simulated Tube Control Polycarbonate can be affected by c...

Page 70: ...roll with optional dispenser and designed for use with Thermo Hybaid s OmniUltra Plate although will give an exceptional seal with other brands of polycarbonate plate TDX Tape Clear in colour and designed for use with polypropylene plates such as OmniTube 96 OmniFast 96 or OmniFast 384 OptiTape and TDX Tape cannot be interchanged as the TDX Tape will bind irreversibly to polycarbonate To secure co...

Page 71: ...ed lid setting Removing the Tape Prior to removing the Tape or OmniSeal Mat from a plate ensure there is no condensation of the solution inside the wells or on the plugs of the tape by either 1 Removing the 96 well plate from the block and cooling to 4 C 2 Cooling down the PCR block to 4 C if possible 3 Heating the sample to 35 C for 30 minutes This will minimise the risk of well to well contamina...

Page 72: ...inutes Sealing a 96 Well Plate OmniSeal TD Mats have 96 plugs in the underside They are spaced so that they fit into the wells of a 96 well plate The plugs are shaped to expand sideways in order to seal all 96 wells of a 96 well plate when sufficient pressure is exerted to the upper surface of the mat by the Px2 heated lid Figure 10 1 Showing how the OmniSeal TD Mat seals the wells of a 96 well pl...

Page 73: ...ally volume loss of 3µl per well in a 30 cycle reaction can be expected The Thermo Hybaid OmniUltra helps to address this problem to some extent In our tests reaction volumes of 10µl gave strong results 5µl reaction volumes gave visible but faint results thus 10µl is the minimum recommended sample volume for this plate This is because this plate is injection moulded which results in absolute well ...

Page 74: ...or probes attached to the surface of slides Optimum results across four standard slides 1mm thickness with the humidity chamber in place have been obtained using a calibration factor of 25 Using a higher calibration factor accelerates the approach to target temperature but results in a degree of overshoot Conversely a lower calibration factor slows the approach to target temperature but minimises ...

Page 75: ...th the Humidity Chamber in place and the lid closed Fill the reservoirs with water Pre heat the block with the Humidity Chamber in place This allows the reservoir to warm up so that evaporation of the solution can begin Alternatively preheat the reservoir solution to the incubation temperature required For thermal cycling with extended high temperature incubations it is important to seal the reage...

Page 76: ...ograms or create extra space for new protocols GRADIENT CALCULATOR Software Software for use in calculating optimised annealing temperatures after gradient cycling HOLD A HOLD step may be specified at the end of a stage Samples are held at this temperature until CONTINUE is pressed to resume the program This function can be used to hold samples at a fixed temperature while a reagent is added befor...

Page 77: ...program on the block Sample Volume When using simulated tube control methods the sample is entered to ensure precise block heating and cooling SET UP This function allows the default settings of the Px2 set day and time power restart option and screen brightness to be modified Simulated Plate Control Simulated Plate Control incorporates an algorithm to reduce time to reach target temperature using...

Page 78: ...10 seconds on a 60 second step will give time intervals of 60 70 80 90 100 seconds This is useful when increasing an incubation step to allow for the depletion of an enzyme Touchdown PCR A gene amplification procedure where the annealing temperature is high e g 65 C to achieve high specificity at the start of the reaction It can then be reduced with successive cycles by e g 0 5 C Cycle to increase...

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Page 80: ...p 3 72 C 0 00 30 x 30 Stage 3 Step 1 72 C 0 05 00 x 1 Hold 4 C PROGRAM F 91 2T_PCR Stage 1 Step 1 94 C 0 01 00 x 1 Stage 2 Step 1 94 C 0 00 10 Step 2 68 C 0 00 10 x 30 Stage 3 Step 1 72 C 0 05 00 x 1 Hold 4 C PROGRAM F 92 LONGPCR Stage 1 Step 1 94 C 0 02 00 x 1 Stage 2 Step 1 94 C 0 00 10 Step 2 65 C 0 00 30 Step 3 68 C 0 10 00 x 10 Stage 3 Step 1 94 C 0 00 10 Step 2 65 C 0 00 30 Step 3 68 C 0 10 ...

Page 81: ...R Stage 1 Step 1 94 C 0 01 00 x 1 Stage 2 Step 1 94 C 0 00 30 Step 2 65 C 0 00 30 Step 3 72 C 0 00 30 x 4 Stage 3 Step 1 94 C 0 00 30 Step 2 65 C 0 00 30 Temp DEC 1 0 C cycle Step 3 72 C 0 00 30 x 16 Stage 4 Step 1 94 C 0 00 30 Step 2 50 C 0 00 30 Step 3 72 C 0 00 30 x 5 Stage 5 Step 1 72 C 0 05 00 x 1 Artisan Technology Group Quality Instrumentation Guaranteed 888 88 SOURCE www artisantg com ...

Page 82: ...AM F 96 RTPCR65 Stage 1 Step 1 65 C 1 00 00 Step 2 94 C 0 01 00 Stage 2 Step 1 94 C 0 00 15 Step 2 65 C 0 00 30 x 40 Stage 3 Step 1 65 C 0 07 00 x 1 PROGRAM F 97 RAMP Stage 1 Step 1 94 C 0 01 00 x 1 Stage 2 Step 1 94 C 0 00 30 Ramp 1 0 C Sec Step2 55 C 0 00 30 Ramp 1 0 C Sec Step 3 72 C 0 00 30 Ramp 1 0 C Sec Stage 3 Step 1 72 C 0 05 00 x 1 Artisan Technology Group Quality Instrumentation Guarante...

Page 83: ... 2002 72 PROGRAM F 98 DIGEST Stage 1 Step 1 37 C 4 00 00 x 1 PROGRAM F 99 CUT KIL Stage 1 Step 1 37 C 4 00 00 Step 2 95 C 0 15 00 x 1 Hold 4 C Artisan Technology Group Quality Instrumentation Guaranteed 888 88 SOURCE www artisantg com ...

Page 84: ...l service in house repair center WE BUY USED EQUIPMENT Sell your excess underutilized and idle used equipment We also offer credit for buy backs and trade ins www artisantg com WeBuyEquipment REMOTE INSPECTION Remotely inspect equipment before purchasing with our interactive website at www instraview com LOOKING FOR MORE INFORMATION Visit us on the web at www artisantg com for more information on ...

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