VTi 90 Rotor - American Instrument Exchange, Inc.
Contents
1. Homogeneous CsCl Solution g mL VTi 90 ROTOR 1 2 20 C 4 C 0 10 20 30 40 50 60 70 80 90 Rotor Speed K rpm Figure 2a Precipitation Curves for the VTi 90 Rotor Using combinations of rotor speeds and homogeneous CsCl solution densities that intersect on or below these curves ensures that CsCl will not precipitate during centrifugation If gradient and sample solutions do not completely fill the tube add mineral oil to fill Do not use an oil overlay in Ultra Clear tubes The dashed line is a representation of equation 6 and is shown here to illustrate the inability of that equation to predict CsCl precipitation 14 VTi 90 Rotor AT EQUILIBRIUM2. During or After Centrifugation Volume mL 19 5 0 4 0 3 0 2 0 1 0 0 1 8 r f 1 7 m q 4 1 6 ws TES OY re S 1 5 or So Sa Q RON J D s Q l a 1 4 Sy Re f Q YF fe P 13 LF 1 2 VTi 90 ROTOR 20 C 4 C 1 1 1 0 1 2 3 4 5 6 7 8 9 10 11 12 13 Distance Across Tube Diameter mm During Centrifugation Figure 2b CsCl Gradients at Equilibrium for the VTi 90 Rotor Centrifugation of homogeneous CsCl solutions at the maximum allowable speeds from Figure 2a results in gradients presented here 15 VTi 90 Rotor AT EQUILIBRIUM During or After Centrifugation Volume mL 19 5 0 4 0 3 0 2 0 1 0 0 AT Or 20 1 8 1 7 ra BP a Le f v zT e o2 pa 1 6 i 14 a a t x T ai 44A o Pa v HT R a j 1 5 7 S a 3 LA ot zi gt Pa 8 Ft ay d a acl 1 4 AE 3 PES Ead Pg HT art mT ma L a ja
3. Arranging Tubes in the Rotor Two four six or eight tubes can be centrifuged per run if they are arranged in the rotor as shown 4 Complete loading by placing the correct spacers and floating spacers if applicable over the tubes It is important that each cavity being used is completely filled 5 Insert a rotor plug 368545 gasket end down over each spacer and screw it in I gt NOTE Do not use rotor plugs in empty cavities 6 Using the hex plug adapter 976959 and torque wrench 858121 tighten each rotor plug to 13 6 Nem 120 in lb To avoid strip ping the plugs apply downward pressure to the hex plug adapter while tightening the plugs Do not overtighten plugs the top surface of each rotor plug should be flush with the surrounding rotor surface OPERATION VTi 90 Rotor Carefully place the rotor on the drive hub Refer to the instrument instruction manual for ultracentrifuge operation For additional operating information see the following e RUN TIMES page 9 for using k factors to adjust run durations e RUN SPEEDS page 11 for information about speed limitations e SLOW ACCELERATION DECELERATION page 11 for infor mation about using slow acceleration and deceleration for gradient stability e SELECTING CsCl GRADIENTS page 13 for methods to avoid CsCl precipitation during centrifugation REMOVAL AND SAMPLE RECOVERY Tube Removal Tool 361668 A CAUTION If disassembly reve
4. China Telephone 86 10 6515 6028 Fax 86 10 6515 6025 6515 6026 CZECH REPUBLIC Beckman Coulter Prague Radiova 1 102 27 Prague 10 Czech Republice Telephone 420 267 00 85 13 Fax 420 267 00 83 23 EASTERN EUROPE MIDDLE EAST NORTH AFRICA Beckman Coulter International S A 22 Rue Juste Olivier Case Postale 301 303 CH 1260 Nyon Switzerland Telephone 41 22 365 3707 Fax 41 22 365 0700 FRANCE Beckman Coulter France S A Paris Nord II 33 66 rue des Vanesses B P 50359 Villepinte France 95942 ROISSY CDG Cedex Telephone 33 1 49 90 90 00 Fax 33 1 49 90 90 10 e mail bioresearch france beckman com GERMANY Beckman Coulter GmbH Europark Fichtenhain B 13 47807 Krefeld Germany Telephone 49 21 51 33 35 Fax 49 21 51 33 33 e mail bioresearch de beckman com GS zraou COULTER Beckman Coulter Inc e Sales and Service 1 800 742 2345 HONG KONG Beckman Coulter Hong Kong Ltd 12th Floor Oxford House 979 King s Road Taikoo Place Hong Kong Telephone 852 2814 7431 Fax 852 2873 4511 INDIA Beckman Coulter India Pvt Ltd Solitaire Corporate Park 3rd Floor Bldg 11 Andheri Ghatkopar Link Road Chakala Andheri West Mumbai India 400 098 Telephone 91 22 3080 5101 ITALY Beckman Coulter S p a Centro Direzionale Lombardo Palazzo F 1 Via Roma 108 20060 Cassina de Pecchi Milano Italy Telephone 39 02 953921 Fax 39 02 95392264 JAPAN Beck
5. 000 462 000 415 000 8 75 000 448 000 406 000 365 000 9 70 000 390 000 354 000 318 000 11 65 000 336 000 305 000 274 000 12 60 000 287 000 260 000 233 000 14 55 000 241 000 219 000 196 000 17 50 000 199 000 181 000 162 000 19 45 000 161 000 146 000 131 000 20 40 000 128 000 116 000 104 000 22 Calculated for all Beckman Coulter preparative rotors as a measure of the rotor s relative efficiency in pelleting sample in water at 20 C Relative Centrifugal Fields for the VTi 90 Rotor 700 000 600 000 500 000 400 000 300 000 200 000 100 000 0 0 10 000 20 000 30 000 40 000 50 000 60 000 70 000 80 000 90 000 Speed rpm 12 VTi 90 Rotor load Note however that the use of this formula may still produce maximum speed figures that are higher than the limitations imposed by the use of certain tubes or adapters In such cases use the lower of the two figures 2 Further speed limits must be imposed when CsCl or other self forming gradient salts are centrifuged as equation 6 does not predict concentration limits speeds that are required to avoid precipitation of salt crystals Precipitation during centrifugation would alter the density distribution of CsCl and this would change the position of the sample bands Figure 2 together with the description and examples below show how to reduce run speeds when using CsCl gradients
6. 90 rotor may void the rotor warranty and should be prohibited by your laboratory safety officer Only the components and accessories listed in this publication should be used in this rotor Read the Safety Notice page at the front of this manual before using the rotor 1 Inspect the rotor plugs and gaskets for damage the high forces generated in this rotor can cause damaged components to fail 2 Make sure that the rotor is equipped with the correct overspeed disk If the disk is missing or damaged replace it according to the instructions in Rotors and Tubes 3 Verify that only the tubes and accessories listed in Table 1 are being used Check the chemical compatibilities of all materials used refer to Appendix A in Rotors and Tubes For runs at other than room temperature refrigerate or warm the rotor beforehand for fast equilibration 1 Be sure that the plug threads are clean and lightly but evenly lubri cated with Spinkote lubricant 306812 to ensure a proper seal by minimizing thread friction 2 Set the rotor in the rotor vise 342705 which should be bolted or clamped to a rigid surface VTi 90 Rotor 3 Load the filled and plugged or sealed tubes symmetrically into the rotor see page 8 for tube information If fewer than eight tubes are being run they must be arranged symmetrically in the rotor see Figure 1 Opposing tubes must be filled to the same level with liquid of the same density Figure 1
7. new gasket snaps onto the grooved end of the plug 18 VTi 90 Rotor Refer to Appendix A in Rotors and Tubes for the chemical resistances of rotor and accessory materials Your Beckman Coulter representa tive provides contact with the Field Rotor Inspection Program and the rotor repair center CLEANING Wash the rotor and rotor components immediately if salts or other corrosive materials are used or if spillage has occurred Do not allow Roir cisarina corrosive materials to dry on the rotor Kit 339558 Under normal use wash the rotor frequently at least weekly to prevent buildup of residues a 1 Wash the rotor plugs and spacers in a mild detergent such as Beckman Solution 555 339555 that won t damage the rotor The Rotor Cleaning Kit 339558 contains two plastic coated brushes and two quarts of Solution 555 for use with rotors and accessories Dilute the detergent 10 to 1 with water gt NOTE Do not wash rotor components in a dishwasher Do not soak in detergent solution for long periods such as overnight E 2 Rinse the cleaned rotor and components with distilled water 3 Air dry the rotor upside down Do not use acetone to dry Gasket the rotor Threads Sk oy Clean plug threads as necessary Use a brush and concentrated NS Solution 555 Rinse and dry thoroughly then lubricate lightly but evenly with Spinkote to coat all threads F 19 VTi 90 Rotor 20 DE
8. not cover the failure of a Beckman Coulter rotor in a centrifuge not of Beckman Coulter manufacture or if the rotor is used in a Beckman Coulter centrifuge that has been modified without the written permission of Beckman Coulter or is used with carriers buckets belts or other devices not of Beckman Coulter manufacture 7 Rotor parts subject to wear including but not limited to rotor O rings VTi NVT TLV MLN and TLN rotor tube cavity plugs and gaskets tubing tools optical overspeed disks bear ings seals and lubrication are excluded from this warranty and should be frequently inspected and replaced if they become worn or damaged 8 Keeping a rotor log is not mandatory but may be desirable for maintenance of good laboratory practices Repair and Replacement Policies 1 Ifa Beckman Coulter rotor is determined by Beckman Coulter to be defective Beckman Coulter will repair or replace it subject to the Warranty Conditions A replacement rotor will be warranted for the time remaining on the original rotor s warranty 2 Ifa Beckman Coulter centrifuge is damaged due to a failure of a rotor covered by this warranty Beckman Coulter will supply free of charge i all centrifuge parts required for repair except the drive unit which will be replaced at the then current price less a credit determined by the total number of revolutions or years completed provided that such a unit was manufactured or rebuilt by Beckman Coulte
9. per run The rotor is made of titanium and is finished with black polyurethane paint A tube spacer and hex cavity rotor plug hold each tube in the rotor and a plug gasket forms a closure around each plug Rotor plugs are black anodized aluminum and spacers are black anodized aluminum for Quick Seal tubes and gold anodized aluminum for OptiSeal tubes Because of the weight of the rotor drive pins are not required in the rotor drive hub cavity For overspeed protection a photoelectric detector in the ultracentri fuge monitors the overspeed disk on the rotor bottom and shuts down the run if speeds exceeding 90 000 rpm are detected See the Warranty at the back of this manual for warranty information PREPARATION AND USE Specific information about the VTi 90 rotor is given here Information common to this and other rotors is contained in Rotors and Tubes for Preparative Ultracentrifuges publication LR IM which should be used together with this manual for complete rotor and accessory operation Publication LR IM is included in the literature package with this rotor manual PRERUN SAFETY CHECKS nN oe 20 Sector gt 355539 nb ROTOR PREPARATION VTi 90 Rotor I gt NOTE Although rotor components and accessories made by other manufacturers may fit in the VTi 90 rotor their safety in this rotor cannot be ascertained by Beckman Coulter Use of other manufacturers components or accessories in the VTi
10. series ultracentrifuges select a slow deceleration profile e in Optima L series ultracentrifuges select SLOW DECEL e in L7 series ultracentrifuges set the brake switch in the 800 rpm position The centrifugal force at a given radius in a rotor is a function of speed Comparisons of forces between different rotors are made by comparing the rotors relative centrifugal fields RCF When rota tional speed is adjusted so that identical samples are subjected to the same RCF in two different rotors the samples are subjected to the same force The RCF at a number of rotor speeds is provided in Table 2 Speeds must be reduced under the following circumstances 1 If nonprecipitating solutions more dense than 1 7 g mL are centri fuged the maximum allowable run speed must be reduced according to the following equation reduced maximum speed 90 000 rpm ji 7 g mL 6 p where p is the density of the tube contents This speed reduction will protect the rotor from excessive stresses due to the added tube 11 VTi 90 Rotor Table 2 Relative Centrifugal Fields for the VTi 90 Rotor 13 5 mL Tubes Entries in this table are calculated from the formula RCF 1 12r RPM 1000 and then rounded to three significant digits Relative Centrifugal Field x g Rotor Speed INC iae At fay INC lial k rpm 71 1 mm 64 5 mm 57 9 mm Factor 90 000 645 000 585 000 525 000 6 85 000 575 000 522 000 469 000 7 80 000 510
11. CONTAMINATION If the rotor or other components are contaminated with toxic or patho genic materials follow appropriate decontamination procedures as outlined by your laboratory safety officer Check Appendix A in Rotors and Tubes to be sure the decontamination method will not damage any part of the rotor STERILIZATION AND DISINFECTION 121 C OO gt Gaon STORAGE e The rotor and all rotor components can be autoclaved at 121 C for up to an hour Remove the plugs from the rotor and place the rotor plugs and spacers in the autoclave upside down e Ethanol 70 or hydrogen peroxide 6 may be used on all rotor components including those made of plastic Bleach sodium hypochlorite may be used but may cause discoloration of anod ized surfaces Use the minimum immersion time for each solution per laboratory standards While Beckman Coulter has tested these methods and found that they do not damage the rotor or components no guarantee of sterility or disinfection is expressed or implied When sterilization or disinfec tion is a concern consult your laboratory safety officer regarding proper methods to use OptiSeal and Quick Seal tubes are disposable and should be discarded after a single use When it is not in use store the rotor in a dry environment not in the instrument with plugs removed to allow air circulation so moisture will not collect in the tube cavities Flammability hazard Do not use in o
12. LXL TB 004D F November 2007 COULTER VTi 90 Rotor Used In Beckman Coulter Class H R and S Preparative Ultracentrifuges 2007 Beckman Coulter Inc Printed in the U S A CA 1107 2 FNG VTi 90 Rotor SAFETY NOTICE This safety notice summarizes information basic to the safe use of the rotor described in this manual The international symbol displayed above is a reminder to the user that all safety instructions should be read and understood before operation or maintenance of this equipment is attempted When you see the symbol on other pages throughout this publication pay special attention to the specific safety information presented Observance of safety precautions will also help to avoid actions that could damage or adversely affect the performance of the rotor This rotor was developed manufactured and tested for safety and reliability as part of a Beckman Coulter ultracentrifuge rotor system Its safety or reliability cannot be assured if used in a centrifuge not of Beckman Coulter s manufacture or in a Beckman Coulter ultracentrifuge that has been modified without Beckman Coulter s approval A SiS i ie Le Handle body fluids with care because they can transmit disease No known test offers complete assurance that such fluids are free of micro organisms Some of the most virulent Hepatitis B and C viruses HIV I V atypical mycobacteria and certain systemic fungi further emphasize the need for aerosol prote
13. SELECTING CsCl GRADIENTS gt NOTE The curves in Figure 2 are for solutions of CsCl salt dissolved in distilled water only If other salts are present in significant concentrations the overall CsCl concentration may need to be reduced Solid CsCl has a density of 4 g mL and if precipitation during centrifugation may cause rotor failure Precipitation will also alter density distribution and therefore sample separation In general lower speeds provide better resolution but longer run times will be required to achieve particle separation and gradient equilibrium Curves are provided up to the maximum rated speed of the rotor Rotor speed is used to control the slope of a CsCl density gradient and must be limited so that CsCl precipitation is avoided Figure 2a gives the CsCl concentration limiting curves for full tubes The refer ence curves in Figure 2b show equilibrium gradients that result from centrifugation using the maximum densities allowed by Figure 2a Each curve in Figure 2b is within the density limits allowed for the VTi 90 rotor each curve was generated for a single run speed using the maximum allowable homogeneous CsCl densities that avoid precipitation at that speed Figure 2c gives the gradients that result from centrifugation using lower than maximum allowable CsCl concentrations These reduced density curves can be used to make particles band more towards the middle of a tube where volume 13 VTi 90 Rotor
14. acity 0 0 kee eee eee 40 8 mL Approximate acceleration time to maximum speed fully loaded v istine 0 0 eee eee eee 9 min Approximate deceleration time from maximum speed fully loaded 0 00 0 eee eee 8 1 2 min Weight of fully loaded rotor 0000 5 5 kg 12 Ib Rotormaterial 5 0 shea lw eee Ad aa aw eis a ee es titanium Relative Centrifugal Field RCF is the ratio of the centrifugal acceleration at a specified radius and speed rw to the standard acceleration of gravity g according to the following formula RCF 72 g where r is the radius in millimeters is the angular velocity in radians per second 2 x RPM 60 and g is the standard acceleration of gravity 9807 mm s2 After substitution RPM 2 RCF 1 12r F000 VTi 90 Rotor DESCRIPTION Plug Gasket 349290 Plug 368545 Rotor Body LE Overspeed Disk 355539 This Beckman Coulter rotor has been manufactured in a registered ISO 9001 or 13485 facility for use with the appropriately classified Beckman Coulter ultracentrifuges The VTi 90 rated for 90 000 rpm is designed to centrifuge up to eight tubes in an upright position Used in Beckman Coulter class H R and S preparative ultracentrifuges the rotor develops centrifugal forces that can efficiently band DNA or isolate proteins on density gradients Up to 40 8 mL of gradient and sample can be centrifuged
15. als evidence of leakage you should assume that some fluid escaped the rotor Apply appropriate decontamination procedures to the centrifuge and accessories Remove the rotor from the centrifuge by lifting it straight up and off the drive hub Return the rotor to the rotor vise Remove the plugs with the torque wrench To avoid stripping the plugs apply downward pressure to the hex plug adapter while loosening the plugs Use the appropriate removal tool see the SUPPLY LIST to remove the spacers and tubes VTi 90 Rotor TUBES 25 C Com gt The VTi 90 rotor uses only OptiSeal and Quick Seal tubes use only the tubes and accessories listed in Table 1 Refer to Rotors and Tubes for information on the chemical resistances of tube and accessory materials OptiSeal and Quick Seal tubes are disposable and should be discarded after a single use Temperature Limits e Plastic tubes have been centrifuge tested for use at temperatures between 2 and 25 C For centrifugation at other temperatures pretest tubes under anticipated run conditions e If tubes are frozen before use make sure that they are thawed to at least 2 C prior to centrifugation Table 1 Available Tubes for the VTi 90 Rotor Use only the items listed here Tube Required Accessory Max Speed Dimensions Part Part RCF and Volume Description Number Description Number Tube Rack k Factor 13 x 51mm Quick Seal 344075 alumi
16. ar velocity of the rotor in radians per second 0 105 x rpm Fmax is the maximum radius and Fmin is the minimum radius After substitution 11 2 533 x10 In inax Fin b rpm Use the k factor in the following equation to estimate the run time t in hours required to pellet particles of known sedimentation coeffi cient s in Svedberg units Run times can be estimated for centrifugation at less than maximum speed by adjusting the k factor as follows 2 7 50 000__ Kaaj au run speed 4 Run times can also be estimated from data established in prior experi ments if the k factor of the previous rotor is known For any two rotors a and b oe 5 For more information on k factors see Use of k Factor for Estimating Run Times from Previously Established Run Conditions publication DS 719 VTi 90 Rotor SLOW ACCELERATION DECELERATION ACCEL PROFILES DECEL PROFILES RUN SPEEDS SPEED RPM RCF Vertical banding of sample and gradient formation occurs with centrifugation With deceleration tube contents reorient back to horizontal position For gradient stability when preformed gradients are used select slow acceleration as follows e in Optima L XP and XL series ultracentrifuges select a slow acceleration profile e in Optima L series ultracentrifuges select SLOW ACCEL For the stability of all gradients during deceleration do the following e in Optima L XP and XL
17. ction Handle other infectious samples according to good laboratory procedures and methods to prevent spread of disease Because spills may generate aerosols observe proper safety precautions for aerosol contain ment Do not run toxic pathogenic or radioactive materials in this rotor without taking appropriate safety precautions Biosafe containment should be used when Risk Group II materials as identified in the World Health Organization Laboratory Biosafety Manual are handled materials of a higher group require more than one level of protection The rotor and accessories are not designed for use with materials capable of developing flammable or explosive vapors Do not centrifuge such materials in nor handle or store them near the ultracentrifuge Although rotor components and accessories made by other manufacturers may fit in the VTi 90 rotor their safety in this rotor cannot be ascertained by Beckman Coulter Use of other manufacturers components or accessories in the VTi 90 rotor may void the rotor warranty and should be prohibited by your laboratory safety officer Only the components and accessories listed in this publication should be used in this rotor Do not run an empty rotor Place filled tubes in at least two opposing cavities Make sure that filled containers are loaded symmetrically into the rotor and that opposing tubes are filled to the same level with liquid of the same density Make sure that cavities in use have the prop
18. ected in Step 2 In this case the 1 6 g mL 70 000 rpm curves in Figure 2c give the best separation These curves intersect the particle buoyant densities in such a way that particles band about 0 5 mm apart at equilibrium during centrifugation lower axis They are separated by about 0 2 mL upper axis Bands are located in the middle of the tube CARE AND MAINTENANCE MAINTENANCE IIIB NOTE T Do not use sharp tools on the rotor that could 831325 cause scratches in the rotor surface Corrosion begins in scratches and may open fissures in the rotor with continued use e Regularly inspect the overspeed disk If it is scratched damaged or missing replace it Replacement instructions are in Rotors and Tubes e Regularly inspect the rotor plugs 368545 for wear worn threads kk a essa will have a shiny appearance Replace worn plugs 349290 NES Check e Regularly lubricate the metal threads in the rotor plugs with a thin nee even coat of Spinkote lubricant Failure to keep these threads lubri cated can result in damaged threads Replace rotor plugs as a set if they show signs of wear 1 e The rotor plug gaskets 349290 require no maintenance except overspeed cleaning Replace damaged gaskets To replace the plug gasket 355539 use the sharpened end of a cotton swab or similar nonmetallic tool to pry the gasket from the plug Do this carefully so that the plug is not damaged The
19. ed problem Use the address label printed on the RGA form when mailing the rotor and or accessories to Beckman Coulter Inc 1050 Page Mill Road Palo Alto CA 94304 Attention Returned Goods Customers located outside the United States should contact their local Beckman Coulter office 21 VTi 90 Rotor 22 SUPPLY LIST I gt NOTE Publications referenced in this manual can be obtained by calling Beckman Coulter at 1 800 742 2345 in the United States or by contacting your local Beckman Coulter office Call Beckman Coulter Sales 1 800 742 2345 in the United States worldwide offices are listed on the back cover of this manual or see the Beckman Coulter Ultracentrifuge Rotors Tubes amp Accessories catalog BR 8101 for detailed information on ordering parts and supplies For your convenience a partial list is given below REPLACEMENT ROTOR PARTS OTHER VTi 90 rotor assembly 2 2 0 0 0 eee eee 356135 Rotor plugs cis ere le ats eee Bleek a let Gaia AOE Boos 368545 Rotor plug gasket 0 eee eee 349290 Overspeed disk 90 000 rpm 2 20 0 000 0 2 eee eee 355539 Rotor vise assembly 0 0 0 e eee eee eee 342705 Tubes and accessories 2 0 cece eee eee eee see Table 1 Quick Seal Cordless Tube Topper kit 60 Hz 358312 Quick Seal Cordless Tube Topper kit 50 Hz Europe 358313 Quick Seal Cordless Tube Topper kit 50 Hz Great Britain 358314 Quick Seal C
20. er spacers inserted before installing the rotor plugs If disassembly reveals evidence of leakage you should assume that some fluid escaped the rotor Apply appropriate decontamination procedures to the centrifuge and accessories if pathogenic or radioactive materials are involved Never exceed the maximum rated speed of the rotor and labware in use Refer to the section on RUN SPEEDS and derate the run speed as appropriate Do not use sharp tools on the rotor that could cause scratches in the rotor surface Corrosion begins in scratches and may open fissures in the rotor with continued use VTi 90 Rotor VTi 90 ROTOR 1 I KA min fav Tmax l ts o Roraton a rete ser SPECIFICATIONS Maximum speed epas denese eA e eee eee ee 90 000 rpm Density rating at maximum speed 0 0 1 7 g mL Relative Centrifugal Field at maximum speed At faa OLIM sola ain die Pea ase 645 000 x g AU Tay 6425 DAM eei ego han eb ieie Pi i aera 585 000 x g Atrain 7 9 MM 3 ects wn Ser ed trea pied s ak 525 000 x g k factor at maximum speed 0 00 c eee ee eee 6 Conditions requiring speed reductions see RUN SPEEDS Number of tube cavities 0 0 ee eee 8 Available tubes 2s 5 8 Sona se Aa ae ae Ree a see Table 1 Nominal tube dimensions largest tube 13 x 51 mm Nominal tube capacity 0 eee eee 5 1 mL Nominal rotor cap
21. icles will band at these locations across the tube diameter lower axis of Figure 2b at equilibrium during centrifugation After centrifugation the bands will reorient top axis If the required gradient curve is not presented in Figure 2b interpo late between the nearest curves and draw it in For example for a run at 75 000 rpm the curve should be drawn between the 70 000 and 80 000 rpm curves The same particles will band along this curve about one half and one third of the way from the right edge of the figure Using the horizontal axis it can be estimated that these parti cles will be about 1 5 mm apart at equilibrium during centrifugation They will be separated by 0 75 mL top axis Example B Knowing particle buoyant densities 1 600 and 1 610 g mL how do you achieve the best separation 1 In Figure 2c sketch in a horizontal line corresponding to each particle s buoyant density 17 VTi 90 Rotor 2 Select the curve at the temperature 20 C that gives the best particle separation Particles will band at points across the tube diameter where the sketched lines intersect this curve lower axis at equilibrium during centrifugation After centrifugation the bands will reorient top axis 3 Note the run speed along the selected curve 4 Select the maximum homogeneous CsCl density that corresponds to the temperature and run speed established above These param eters will provide the particle banding pattern sel
22. is valid for the time periods indicated above from the date of shipment to the original Buyer by Beckman Coulter or an authorized Beckman Coulter representative 2 This warranty extends only to the original Buyer and may not be assigned or extended to a third person without written consent of Beckman Coulter 3 This warranty covers the Beckman Coulter Centrifuge Systems only including but not limited to the centrifuge rotor and accessories and Beckman Coulter shall not be liable for damage to or loss of the user s sample non Beckman Coulter tubes adapters or other rotor contents 4 This warranty is void if the Beckman Coulter Centrifuge Sys tem is determined by Beckman Coulter to have been operated or maintained in a manner contrary to the instructions in the operator s manual s for the Beckman Coulter Centrifuge System components in use This includes but is not limited to operator misuse abuse or negligence regarding indicated main tenance procedures centrifuge and rotor classification require ments proper speed reduction for the high density of certain fluids tubes and tube caps speed reduction for precipitating gradient materials and speed reduction for high temperature operation 5 Rotor bucket sets purchased concurrently with or subsequent to the purchase of a Swinging Bucket Rotor are warranted only for a term co extensive with that of the rotor for which the bucket sets are purchased 6 This warranty does
23. l 13 AZ oi h pe a Le Hrt Z PES 1 2 VTi 90 ROTOR 90 000 80000 70000 4 60000 1 0 1 2 3 4 5 6 7 8 9 10 11 12 13 Distance Across Tube Diameter mm During Centrifugation Figure 2c CsCl Gradients for Lower Densities Densities used to generate curves are printed along the curves Note that at 4 C only three curves 60 000 rpm 70 000 rpm and 80 000 rpm are shown for 1 6 g mL 16 because CsCl would precipitate at higher speeds VTi 90 Rotor between bands will be greatest The gradients in Figures 2b and 2c can be generated from step or linear gradients or from homogeneous solutions But the total amount of CsCl in solution must be equivalent to a homogeneous solution corresponding to the concentrations specified TYPICAL EXAMPLES FOR DETERMINING CsCl RUN PARAMETERS Example A Knowing homogeneous CsCl solution density 1 64 g mL and approximate particle buoyant densities 1 70 and 1 65 g mL at 20 C where will particles band 1 In Figure 2a find the curve that corresponds to the required run temperature 20 C The maximum allowable rotor speed is deter mined from the point where this curve intersects the homogeneous CsCl density 80 000 rpm 2 In Figure 2b sketch in a horizontal line corresponding to each particle s buoyant density 3 Mark the point in the figure where each particle density intersects the curve corresponding to the selected run speed and temperature 4 Part
24. man Coulter K K TOC Ariake West Tower 2 5 7 Ariake Koto ku Tokyo 135 0063 Japan Telephone 81 3 5530 8500 Fax 81 3 5404 8436 MEXICO Beckman Coulter de Mexico S A de C V Avenida Popocat petl 396 Colonia Gral Pedro Maria Anaya Codigo Postal 03340 Mexico D F Mexico Telephone 001 52 55 9183 2800 NETHERLANDS Beckman Coulter Nederland B V Nijverheidsweg 21 3641 RP Mijdrecht Postbus 47 3640 AA Mijdrecht The Netherlands Telephone 31 297 230630 Fax 31 297 288082 SINGAPORE Beckman Coulter Singapore Pte Ltd 116 Changi Road Unit 03 01 02 Singapore 419718 Telephone 65 6339 3633 Fax 65 6336 6303 SOUTH AFRICA SUB SAHARAN AFRICA Beckman Coulter Stand 1A Primegro Park Tonetti Street 1685 Halfway House Johannesburg Republic of South Africa Telephone 27 11 805 2014 5 Fax 27 11 805 4120 e mail beckman intekom co za SPAIN Beckman Coulter Espafia S A C Caleruega 81 28033 Madrid Spain Telephone 34 91 3836080 Fax 34 91 3836096 email bioresearch espana beckman com SWEDEN Beckman Coulter AB Archimedesvaegen 7 Box 111 56 SE 168 11 Bromma Sweden Telephone 46 8 564 85 900 Telefax 46 8 564 85 901 SWITZERLAND Beckman Coulter International S A 22 Rue Juste Olivier Case Postale 301 303 CH 1260 Nyon Switzerland Telephone 41 0800 850 810 Fax 41 0848 850 810 TAIWAN Beckman Coulter Taiwan Inc Taiwan Branch 8th Floor 216 Tu
25. n Hwa South Road Section 2 Taipei 106 Taiwan Republic of China Telephone 886 2 2378 3456 Fax 886 2 2377 0408 TURKEY Beckman Coulter Ltd E 5 Yanyol Faith Cad 81410 Soganlik Kartal Istanbul Turkey Telephone 90 216 309 1900 Fax 90 216 309 0090 UNITED KINGDOM Beckman Coulter United Kingdom Ltd Oakley Court Kingsmead Business Park London Road High Wycombe Buckinghamshire HP11 1JU England U K Telephone 44 01494 441181 Fax 44 01494 447558 e mail beckman_uk beckman com Additional addresses are available at www beckmancoulter com 4300 N Harbor Boulevard Box 3100 Fullerton California 92834 3100 Internet www beckmancoulter com Telex 678413 Fax 1 800 643 4366 2007 Beckman Coulter Inc G Bd Printed on recycled paper
26. num spacer 342883 348122 90 000 rpm 5 1 mL Ultra Clear pkg 50 645 000 x g 6 13x51mm_ Quick Seal 342412 aluminum spacer 342883 348122 90 000 rom 5 1 mL polyallomer pkg 50 645 000 x g 6 13 x 51 mm OptiSeal 362185 gold anodized 362198 360534 90 000 rom 4 9 mL polyallomer pkg 56 aluminum spacer 645 000 x g 6 13 x 31 5 mm Quick Seal 349621 aluminum spacer 342883 348122 90 000 rom 3 5 mL polyallomer pkg 50 645 000 x g floating spacer 356866 6 13x 25mm Quick Seal 345829 aluminum spacer 342883 348122 90 000 rem 2mL polyallomer pkg 50 645 000 x g floating spacer 345827 6 Includes disposable plastic plugs t Floating spacers part of the g Max system of tube support are made of Noryl a registered trademark of GE Plastics Gold Aluminum _ Spacer S Plug Aluminum Spacer N oy Dome Pa shaped Tube QS Metal A Space I Floating Spacer Bell Top A abe KK RUN TIMES TIME HR MIN a Pe Les l al VTi 90 Rotor OptiSeal Tubes OptiSeal tubes come with plastic plugs and can be quickly and easily prepared for use without tools or heat With the tube spacer and rotor plug in place the combination of g force and hydrostatic pressure during centrifugation ensures a tight reliable seal that protects your samples Fill each tube to the base of the stem leaving no fluid in the stem Overfilling the tube can cause spillage when the plug is inserted o
27. ordless Tube Topper kit 50 Hz Australia 358315 Quick Seal Cordless Tube Topper kit 50 Hz Canada 367803 Tube Topper rack 13 mm dia tubes 004 348122 TOrque Wrens sss miraci Ss ee od WE Be EE ae Se 858121 Hex plug adapter 1 0 ee eee eee 976959 Tube removal tool 0 ec eee cece eee eens 361668 Floating spacer removal tool 0 0 00 02 eee eee eee 338765 Spinkote lubricant 1 0Z 0 0 0 cece eee eee 306812 Silicone vacuum grease 2 OZ 2 2 eee eee eee eee 335148 Rotor Cleaning Kitr er aerosani EEA E E EEEE ee 339558 Beckman Solution 555 1 qt 2 2 ee eee eee eee 339555 Rotor cleaning brush seses 00 0 pi eee eee eee 339379 ULTRACENTRIFUGE ROTOR WARRANTY All Beckman Coulter ultracentrifuge Fixed Angle Vertical Tube Near Vertical Tube Swinging Bucket and Airfuge rotors are warranted against defects in materials or workmanship for the time periods indicated below subject to the Warranty Conditions stated below Preparative Ultracentrifuge Rotors 5 years No Proration Analytical Ultracentrifuge Rotors 5 years No Proration ML and TL Series Ultracentrifuge ROOTS eaen 0 esata e aon Cais Ree 5 years No Proration Airfuge Ultracentrifuge Rotors 1 year No Proration For Zonal Continuous Flow Component Test and Rock Core ultracentrifuge rotors see separate warranty Warranty Conditions as applicable 1 This warranty
28. r and ii if the centrifuge is cur rently covered by a Beckman Coulter warranty or Full Service Agreement all labor necessary for repair of the centrifuge 3 If a Beckman Coulter rotor covered by this warranty is dam aged due to a malfunction of a Beckman Coulter ultracentrifuge covered by an Ultracentrifuge System Service Agreement Beckman Coulter will repair or replace the rotor free of charge 4 If a Beckman Coulter rotor covered by this warranty is damaged due to a failure of a Beckman Coulter tube bottle tube cap spacer or adapter covered under the Conditions of this Warranty Beckman Coulter will repair or replace the rotor and repair the instrument as per the conditions in policy point 2 above and the replacement policy 5 Damage to a Beckman Coulter rotor or instrument due to the failure or malfunction of a non Beckman Coulter tube bottle tube cap spacer or adapter is not covered under this warranty although Beckman Coulter will assist in seeking compensation under the manufacturer s warranty Disclaimer IT IS EXPRESSLY AGREED THAT THE ABOVE WARRANTY SHALL BE IN LIEU OF ALL WARRANTIES OF FITNESS AND OF THE WARRANTY OF MERCHANTABILITY AND BECKMAN COULTER INC SHALL HAVE NO LIABILITY FOR SPECIAL OR CONSEQUENTIAL DAMAGES OF ANY KIND WHATSOEVER ARISING OUT OF THE MANUFAC TURE USE SALE HANDLING REPAIR MAINTENANCE OR REPLACEMENT OF THE PRODUCT Factory Rotor Inspection Service Beckman Coulter Inc
29. r compromise seal integrity however too much air can cause the tube to deform disrupting gradients and sample bands Refer to Using OptiSeal Tubes publication IN 189 included in each package of OptiSeal tubes for detailed information on the use and care of OptiSeal tubes Quick Seal Tubes Quick Seal tubes must be sealed prior to centrifugation These tubes are heat sealed and do not need caps however spacers are required on top of the tubes when they are loaded into the rotor e Fill Quick Seal tubes leaving a small bubble of air at the base of the neck Do not leave a large air space too much air can cause excessive tube deformation and make the tube difficult to remove e Some of the Quick Seal tubes listed in Table 1 are part of the g Max system which uses a combination of small bell top Quick Seal tubes and floating spacers also called g Max spacers This means that you can run the shorter tubes in this rotor without reduction in g force For detailed information on the g Max system see publication DS 709 Refer to Rotors and Tubes for detailed information on the use of Quick Seal tubes The k factor of the rotor is a measure of the rotor s pelleting efficiency Beckman Coulter has calculated the k factors for all of its preparative rotors at maximum rated speed and using full tubes The k factor is calculated from the formula VTi 90 Rotor 10 ie In 7max min 19 x 1 o2 3600 where wis the angul
30. r near operating ultracentrifuges VTi 90 Rotor RETURNING A ROTOR Before returning a rotor or accessory for any reason prior permission RGA a Returned Goods Authorization form must be obtained from Beckman Coulter Inc This RGA form may be obtained from your local Beckman Coulter sales office and should contain the following r information serial number e history of use approximate frequency of use e reason for the return e original purchase order number billing number and shipping number if possible e name and phone number of the person to be notified upon receipt of the rotor or accessory at the factory e name and phone number of the person to be notified about repair costs etc To protect our personnel it is the customer s responsibility to ensure that all parts are free from pathogens and or radioactivity Steriliza tion and decontamination must be done before returning the parts Smaller items such as tubes bottles etc should be enclosed in a sealed plastic bag All parts must be accompanied by a note plainly visible on the out side of the box or bag stating that they are safe to handle and that they are not contaminated with pathogens or radioactivity Failure to attach this notification will result in return or disposal of the items without review of the report
31. will provide free mechanical and metallurgical inspection in Palo Alto California USA of any Beckman Coulter rotor at the request of the user Shipping charges to Beckman Coulter are the responsibility of the user Rotors will be inspected in the user s laboratory if the centrifuge in which they are used is covered by an appropriate Beckman Coulter Service Agreement Contact your local Beckman Coulter office for details of service coverage or cost Before shipping contact the nearest Beckman Coulter Sales and Service office and request a Returned Goods Authorization RGA form and packaging instructions Please include the complete rotor assembly with buckets lid handle tube cavity caps etc A SIGNED STATEMENT THAT THE ROTOR AND ACCESSO RIES ARE NON RADIOACTIVE NON PATHOGENIC NON TOXIC AND OTHERWISE SAFE TO SHIP AND HANDLE IS REQUIRED Beckman Coulter Worldwide Biomedical Research Division Offices AUSTRALIA Beckman Coulter Australia Pty Ltd Unit D 24 College St Gladesville NSW 2111 Australia Telephone 61 2 9844 6000 or toll free 1 800 060 880 Fax 61 2 9844 6096 email lifescienceaustralia beckman com CANADA Beckman Coulter Canada Inc 6755 Mississauga Road Suite 600 Mississauga Ontario Canada L5N 7Y2 Telephone 1 905 819 1234 Fax 1 905 819 1485 CHINA Beckman Coulter Inc Beijing Representative Office Unit 2005A 2006 2009 East Ocean Center Jian Guomenwai Avenue Beijing 100004
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