TLA-120.2 Rotor
Contents
1. 3 4 Ful 1 2 Full 1 4 Full 27 4 30 0 32 5 1 9 1 8 DA j p A m i 17 go 000 P 60 000 P Kj idi i o rem 10 00 1 6 9 re 20 00 m 400 vor g o NS 15 oo 995 go 9S s a xe X Sq eo AQ AO 14 T 1 Xe e SA goo NS ANO AN M NM BAS Fo x TLA 120 2 ROTOR 12 20 C 4 C 11 1 0 21 4 23 5 25 6 27 7 29 8 31 9 34 1 36 2 38 3 min Distance from Axis of Rotation mm max Figure 4 CsCl Gradients at Equilibrium Centrifugation of homogeneous CsCl solutions at the maximum allowable speeds from Figure 3 results in gradients presented here 15 TLA 120 2 Rotor TYPICAL EXAMPLES FOR DETERMINING CsCl RUN PARAMETERS At Speed Floating Components E Bands Pelleted Material At Rest in Rotor Upright 16 Example A A separation that is done frequently is the banding of plasmid DNA in cesium chloride with ethidium bromide The starting density of the CsCl solution is 1 55 g mL In this separation the covalently closed circular plasmid bands at a density of 1 57 g mL while the nicked and linear species band at 1 53 g mL At 20 C where will particles band 1 In Figure 3 find the curve that corresponds to the required run temperature 20 C and fill volume full The maximum allow able rotor speed is determined from the point where this curve interse2. For runs at other than room temperature refrigerate or warm the rotor beforehand for fast equilibration 1 Lightly but evenly lubricate metal threads with Spinkote lubricant 306812 Apply a thin film of silicone vacuum grease 335148 to the two O rings in the rotor lid Load the filled and sealed if required tubes symmetrically into the rotor see page 8 for tube information If fewer than ten tubes are being run they must be arranged symmetrically in the rotor see Figure 2 Opposing tubes must be filled to the same level with liquid of the same density Use the required spacers if necessary see Table 1 to complete the loading operation After the rotor is loaded insert it into the portable polypropylene rotor vise 346133 Place the lid on the rotor and tighten it firmly to the right clockwise by hand No tool is required Figure 2 Arranging Tubes in the Rotor Two four five six eight or ten tubes can be centrifuged per run if they are arranged in the rotor as shown Use an absorbent towel to wipe off condensation from the rotor then carefully place the rotor on the drive hub Lock the rotor in place by gently pressing the plunger down until you feel it click When you remove your finger the plunger will remain flush with the rotor body if it is properly engaged If the plunger pops up repeat the procedure The Optima MAX MAX XP and MAX E ultracentrifuges
3. 348299 CentriTube Slicer adapter for 13 mm tubes 354526 Spinkote lubricant 2 oZ 0 0 eee ete eee 306812 Silicone vacuum grease 102 0 0 cece eee eee ee 335148 Rotor Cleaning Kit ces ieee ies cet iets be eae ee 339558 Rotor cleaning brush 0 0 0 eee eee eee eee 347404 Beckman Solution 555 1 qt 0 cee ee eee eee 339555 21 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 sic omues oed ates Sees 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 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 w
4. RUN TIMES TIME HR MIN usu TLA 120 2 Rotor 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 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 Refer to Rotors and Tubes for detailed information on the use and care of Quick Seal tubes Thickwall Tubes Thickwall polyallomer and polycarbonate tubes can be run partially filled at least half filled without caps but all opposing tubes for a run must be filled to the same level with liquid of the same density Do not overfill capless tubes be sure to note the reduction in run speed shown in Table 1 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 In max min 19 x 1 d 3600 where cis the angular velocity of the rotor in radians per second 0 105 x rpm Fmax is the maximum radius and rq is the minimum radius TLA 120 2 Rotor 10 RUN SPEEDS SPEED RPM RCF After substitution rpm 2 Use the k factor in the following equation to estimate the run time t in hou
5. 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 1 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 Coulter 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
6. 339555 that won t damage the rotor The Rotor Cleaning Kit 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 III NOTE Do not wash rotor components in a dishwasher Do not soak in detergent solution for long periods such as overnight 3 Rinse the cleaned rotor and components with distilled water 4 Air dry the rotor and lid upside down Do not use acetone to dry the rotor 5 Apply a thin even coat of silicone vacuum grease to both lid O rings before replacing them in the grooves in the lid Clean metal threads as necessary at least every 6 months Use a brush and concentrated Solution 555 Rinse and dry thoroughly then lubricate lightly but evenly with Spinkote to coat all threads Periodically remove the O rings and wipe clean as necessary Clean the O ring grooves with a cotton tipped swab Reapply a light film of silicone vacuum grease If the rotor or other components are contaminated with radioactive toxic or pathogenic materials follow appropriate decontamination procedures as outlined by appropriate laboratory safety guidelines and or other regulations Consult Appendix A in Rotors and Tubes to select an agent that will not damage the rotor TLA 120 2 Rotor STERILIZATION AND DISINFECTION The rotor and all rotor components can be autoclaved at 121 C for up to an hour Remove the lid and O rings from the rotor
7. 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 Fax 1 800 643 4366 CA Bo Printed on recycled paper
8. 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 C COULTER Beckman Coulter Inc 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 Beckman 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 NETHER
9. automatically secures the rotor to the drive shaft without the need for engaging the plunger TLA 120 2 Rotor A CAUTION In all ultracentrifuge models except the Optima MAX MAX XP and MAX E it is very important to lock the rotor in place before begin ning the run to ensure that the rotor remains seated during centrifugation Failure to lock the rotor in place before beginning the run may result in damage to both rotor and instrument 3 Refer to the instrument instruction manual for ultracentrifuge operation 4 For additional operating information see the following RUN TIMES page 9 for using k factors to adjust run durations RUN SPEEDS page 10 for information about speed limitations SELECTING CsCl GRADIENTS page 12 for methods to avoid CsCl precipitation during centrifugation REMOVAL AND SAMPLE RECOVERY Plunger Released N CAUTION If disassembly reveals evidence of leakage you should assume that some fluid escaped the rotor Apply appropriate decontamination procedures to the centrifuge and accessories 1 To release the plunger at the end of the run gently press it down until you feel it click When you remove your finger the plunger will pop up to its released position 2 Remove the rotor from the ultracentrifuge and place it in the rotor vise 3 Remove the lid by unscrewing it to the left counterclockwise 4 Use a tube removal tool to remove the
10. spacers and tubes TLA 120 2 Rotor TUBES AND ACCESSORIES The TLA 120 2 rotor uses tubes and accessories listed in Table 1 Be sure to use only those items listed and to observe the maximum speed limits shown Refer to Appendix A in Rotors and Tubes for information on the chemical resistances of tube and accessory materials Table 1 Available Tubes for the TLA 120 2 Rotor Use only the items listed here and observe maximum fill volumes and speeds shown Tube Required Accessory Max Speed Dimensions Part Part RCF and Volume Description Number Description Number k Factor 11 x 34 mm thickwall 343778 none 120 000 rpm 1 0 mL polycarbonate pkg 100 627 000 x g 8 11 x 34mm thickwall 347287 none 80 000 rpm 1 0 mL polyallomer pkg 100 279 000 x g 18 11 x 25 mm Quick Seal 344624 Noryl spacer 344636 120 000 rpm 1 5 mL polyallomer pkg 50 pkg 6 627 000 x g 14 11 x 32 mm Quick Seal 344625 Noryl spacer 344674 120 000 rpm 2 0 mL polyallomer pkg 50 pkg 6 627 000 x g 16 N a o O O gt Noryl is a registered trademark of GE Plastics Temperature Limits 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 f plastic containers are frozen before use make sure that they are thawed to at least 2 C prior to centrifugation Spacer D
11. 480 000 393 000 303 000 11 100 000 436 000 356 000 274 000 12 95 000 393 000 321 000 248 000 13 90 000 353 000 288 000 222 000 14 85 000 315 000 257 000 198 000 16 80 000 279 000 228 000 176 000 18 75 000 245 000 200 000 154 000 21 70 000 213 000 175 000 134 000 24 65 000 184 000 150 000 116 000 28 60 000 157 000 128 000 98 800 33 55 000 132 000 108 000 83 000 39 50 000 109 000 89 000 68 600 47 Calculated for all Beckman Coulter preparative rotors as a measure of the rotor s relative pelleting efficiency in water at 20 C Relative Centrifugal Fields for the TLA 120 2 Rotor 60 0000 50 0000 40 0000 30 0000 20 0000 10 0000 20 000 40 000 60 000 Speed rpm 80 000 100 000 120 000 TLA 120 2 Rotor 11 TLA 120 2 Rotor 12 Speeds must be reduced under the following circumstances 1 If nonprecipitating solutions more dense than 1 7 g mL are centrifuged the maximum allowable run speed must be reduced according to the following equation reduced maximum speed 120 000 rpm 17 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 load Note however that the use of this formula may still produce maximum speed values that are higher than the limitations imposed by the use of certain tubes or adapters In such cases use the lower of the two v
12. LANDS 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 Internet www beckmancoulter com Telex 678413 2007 Beckman Coulter Inc 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 Tun 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
13. 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 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 a
14. TL TB 016G October 2007 COULTER TLA 120 2 Rotor Used In Beckman Coulter Optima MAX XP MAX MAX E TL TLX and TL 100 Tabletop Ultracentrifuges 2007 Beckman Coulter Inc Printed in the U S A TLA 120 2 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 informa tion 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 E ps pepe S 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 empha
15. alues 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 Figures 3 and 4 together with the description and examples below show how to reduce run speeds when using CsCl gradients SELECTING CsCl GRADIENTS Precipitation during centrifugation would alter density distribution and this would change the position of the sample bands Curves in Figures 3 and 4 are provided up to the maximum rated speed of the rotor Curves in Figures 3 and 4 were generated using the 1 5 mL Quick Seal tube gt NOTE The curves in Figures 3 and 4 are for solutions of CsCl salt dissolved in distilled water only If other salts are present in significant concentra tions the overall CsCl concentration may need to be reduced ADJUSTING FILL VOLUMES TLA 120 2 Rotor Rotor speed is used to control the slope of a CsCl density gradient and must be limited so that CsCl precipitation is avoided Speed and density combinations that intersect on or below the curves in Figure 3 ensure that CsCl will not precipitate during centrifugation in the TLA 120 2 rotor Curves are provided at two temperatures 20 C black curves and 4 C gray curves The ref
16. and place TED the rotor and lid in the autoclave upside down m Ethanol 7096 or hydrogen peroxide 696 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 Guu 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 STORAGE When it is not in use store the rotor in a dry environment not in the instrument with the lid removed to allow air circulation so moisture will not collect in the tube cavities Flammability hazard Do not use in or near operating ultracentrifuges 19 TLA 120 2 Rotor 20 RETURNING A ROTOR Before returning a rotor or accessory for any reason prior permission 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 information serial number history of use approximate frequency of use reason for the return original purchase order number billing
17. arranty 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 maintenance procedures centrifuge and rotor classification requirements proper speed reduction for the high density of certain fluids tubes and tube caps speed reduction for precipi tating gradient materials and speed reduction for high tempera ture 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 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
18. at 84 000 rpm Figure 5 presents the gradient profile use the three quarter filled segment only A tube full of the 1 70 g mL CsCl solution may be centrifuged no faster than 79 000 rpm 13 TLA 120 2 Rotor 14 Homogeneous CsCl Solution g mL 1 9 1 8 S iL L L3 o Af x 1 L5 Pag 2 1 1 6 KA ZAREN 1 5 TLA 120 2 ROTOR 1 4 20 C 4 C 1 3 1 2 1 1 0 10 20 30 40 50 60 70 80 90 100 110 120 Rotor Speed K rpm Figure 3 Precipitation Curves for the TLA 120 2 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 Density g mL TLA 120 2 Rotor
19. back cover of this manual or see the Beckman Coulter Ultracentrifuge Rotors Tubes amp Acces sories catalog BR 8101 available at www beckmancoulter com for detailed information on ordering parts and supplies For your conve nience a partial list is given below REPLACEMENT ROTOR PARTS OTHER TLA 120 2 rotor assembly 00 0 0 eee eee eee eee 362046 Lidvassembly scant wt Pe NP Ba he Ss coe dx pcd 362048 Ed Oting outer ose Hee che S CES RESP RIDERE 854519 Lid O ring inner i oteemtbemebv a eb b e Wesen va 824412 ROOT VISE 2 cns eet eR DREDRUNDOE DEEP OPE TN B Eus 346133 Tubes and accessories n 0 ccc eee ees see Table 1 TUDE TACK icce Sete eee EE e See ree tese 349387 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 Cordless Tube Topper kit 50 Hz Australia 358315 Quick Seal Cordless Tube Topper kit 50 Hz Canada 367803 Tube removal tool Aesir ereen tote a en e eee 361668 Curved hemostat 6 iN 0 0 0 cece eee eene 927208 Straight hemostat 6 1n 2 eee eee 961519 Fraction Recovery System 0 0 0 cece eee eee 342025 Fraction Recovery System Adapter Kit for TL series tubes 347828 Beckman Coulter CentriTube Slicer 00005 347960 CentriTube Slicer replacement blades pkg of 10
20. cts the homogeneous CsCl density 110 000 rpm 2 In Figure 4 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 Particles will band at these locations across the tube diameter at equilibrium during centrifugation In this example particles will band about 32 1 and 30 0 mm from the tube bottom about 2 1 mm of centerband to centerband separation at the rotor s 30 degree tube angle When the tube is held upright there will be about 2 4 mm of centerband to centerband separation This interband distance dup can be calculated from the formula d 7 d UP cos 0 where d is the interband distance when the tube is held at an angle 0 in the rotor Example B Knowing particle buoyant densities for example 1 66 and 1 62 g mL how do you achieve good separation using open top tubes 1 In Figure 4 sketch in a horizontal line corresponding to each particle s buoyant density 2 Select the curve at the required temperature 20 C and fill volume that gives the best particle separation 3 Note the run speed along the selected curve 80 000 rpm TLA 120 2 Rotor 4 From Figure 4 select the maximum homogeneous CsCl density that corresponds to the temperature and run speed established above These parameters will provide the particle bandi
21. erence curves in Figure 4 show gradient distribution at equi librium Each curve in Figure 4 is within the density limits allowed for the TLA 120 2 rotor each curve was generated for a single run speed using the maximum allowable homogeneous CsCl densities one for each fill level that avoid precipitation at that speed The gradients in Figure 4 can be generated from step or linear gradients or from homogeneous solutions But the total amount of CsCl in solu tion must be equivalent to a homogeneous solution corresponding to the concentrations specified in Figure 3 Figure 4 can also be used to approximate the banding positions of sample particles Figures 3 and 4 show that several fill volumes are possible in a tube If a tube is partially filled with gradient solution float mineral oil or some other low density immiscible liquid on top of the tube contents to fill the tube to its maximum volume Note that for a given CsCl density as the fill level decreases the maximum allowable speed increases Partial filling may be desirable when there is little sample or when you wish to shorten the run time For example a one quarter filled tube of a 1 70 g mL homogeneous CsCl solution at 20 C may be centrifuged at 105 000 rpm see Figure 4 The segment of the 105 000 rpm curve Figure 5 from the one quarter filled line to 1 86 g mL at the tube bottom represents this gradient The same solution in a three quarter filled tube may be centrifuged
22. fore installing the rotor lid If disassembly reveals evidence of leakage you should assume that some fluid escaped the rotor Apply appropriate decontamination procedures to the centrifuge and accessories 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 TLA 120 2 ROTOR SPECIFICATIONS TLA 120 2 Rotor od I min _ A U S Pat No 4 372 483 max Japanese Pat No 1 551 443 Swiss Pat No 646 881 Axis of Rotation Maximum speed 00 0 e eee eee eee eee 120 000 rpm Density rating at maximum speed 000 1 7 g mL Relative Centrifugal Field at maximum speed Ata OS O mni 2454 Aas eee eae Aken kate PRO 627 000 x g Atay B18 MM RR ERE EP E RP RR 513 000 x g ACh 24 50 inire ee x ER E Ue quA 395 000 x g k factor at maximum speed 0 2 eee eee eee 8 Conditions requiring speed reductions see RUN SPEEDS Number of tube cavities 2 0 eee eee 10 Available tubes 0 0 eee eee eee see Table 1 Nominal tube dimensions largest tube 11 x 34 mm Nominal tube capacity open top sssse
23. n 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 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 CHINA Beckman Coulter Inc Beijing Representative Office Unit 2005A 2006 2009 East Ocean Center Jian Guomenwai Avenue Beijing 100004 China Telephone 86 10 6515 6028 Fax 86 10 6515 6025 6515 6026 EASTERN EUROPE MIDDLE EAST NORTH AFRICA Beckman Coulter International S A 22 Rue Juste Olivier Case
24. ng pattern selected in Step 2 CARE AND MAINTENANCE MAINTENANCE gt NOTE 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 Lid pen IN e Regularly lubricate the metal threads in the rotor with a thin even Le coat of Spinkote lubricant Failure to keep these threads lubricated Lid O ring can result in damaged threads outer Lid 854513 oxing Regularly apply silicone vacuum grease to the O rings Replace i 2 Sheck 8244 12 O rings about twice a year or whenever worn or damaged for Corrosion y D CLEANING Rotor Cleaning Kit 339558 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 Wash the rotor and rotor components immediately if salts or other corrosive materials are used or if spillage has occurred Do not allow corrosive materials to dry on the rotor Under normal use wash the rotor frequently at least weekly to prevent buildup of residues 17 TLA 120 2 Rotor 18 outer 854519 x Lid O ring inner iz 824412 DECONTAMINATION 1 Remove the O rings before washing 2 Wash the rotor and lid in a mild detergent such as Beckman Solution 555
25. number and shipping number if possible name and phone number of the person to be notified upon receipt of the rotor or accessory at the factory 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 reported 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 SUPPLY LIST TLA 120 2 Rotor 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 Contact Beckman Coulter Sales 1 800 742 2345 in the United States worldwide offices are listed on the
26. rs required to pellet particles of known sedimentation coeffi cient s in Svedberg units 5 t 3 Run times can be estimated for centrifugation at less than maximum speed by adjusting the k factor as follows kay t L0 a adj ctual run speed Run times can also be estimated from data established in prior experiments if the k factor of the previous rotor is known For any two rotors a and b t k 4 5 fp b For more information on k factors see Use of k Factor for Estimating Run Times from Previously Established Run Conditions publication DS 719 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 rotational 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 RCF x g 70 0000 Table 2 Relative Centrifugal Fields for the TLA 120 2 Rotor 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 At Imax At ray At fmin k rpm 38 9 mm 31 8 mm 24 5 mm Factor 120 000 627 000 513 000 395 000 8 115 000 576 000 471 000 363 000 9 110 000 527 000 431 000 332 000 10 105 000
27. ses esses 1 0 mL Nominal tube capacity Quick Seal 0 005 2 0 mL Nominal rotor capacity 0 2 eee eee 20 mL Approximate acceleration time to maximum speed fully loaded 2 0 0 0 eee eee eee 2 1 2 min Approximate deceleration time from maximum speed fully loaded 0 0 eee eee ee eee 2 min Weight of fully loaded rotor 000 0 6 kg 1 2 Ib Rotor material cues RR quc edie Seige d donc ed titanium Relative Centrifugal Field RCF is the ratio of the centrifugal acceleration at a specified radius and speed ro to the standard acceleration of gravity g according to the following formula RCF 222 8 where r is the radius in millimeters is the angular velocity in radians per second 2 z RPM 60 and g is the standard acceleration of gravity 9807 mm s2 After substitution RPM 1000 RCF 1 12r TLA 120 2 Rotor DESCRIPTION w Lid Assembly 362048 Lid O ring outer 854519 Lid O ring inner 824412 ON Rotor Body 362046 This rotor has been manufactured in a registered ISO 9001 or 13485 facility for use with the specified Beckman Coulter ultracentrifuges The TLA 120 2 fixed angle rotor rated for 120 000 rpm has a tube angle of 30 degrees from the axis of rotation The rotor can centrifuge up to 10 tubes and is used in Beckman Coulter Optima MAX MAX E MAX XP TL TLX and the TL 100 tabletop ultracen
28. size the need for aerosol protection 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 TLA 120 2 rotor their safety in this rotor cannot be ascertained by Beckman Coulter Use of other manufacturers components or accessories in the TLA 120 2 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 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 proper spacers and or floating spacers inserted be
29. t exceed its maximum permitted speed See the Warranty at the back of this manual for warranty information PREPARATION AND USE Specific information about the TLA 120 2 rotor is given here Information common to this and other rotors is contained in Rotors and Tubes for Tabletop Preparative Ultracentrifuges publication TLR IM which should be used together with this manual for complete rotor and accessory operation Publication TLR IM is included in the literature package shipped with the rotor gt NOTE Although rotor components and accessories made by other manufacturers may fit in the TLA 120 2 rotor their safety in this rotor cannot be ascertained by Beckman Coulter Use of other manufacturers components or accessories in the TLA 120 2 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 PRERUN SAFETY CHECKS Read the Safety Notice page at the front of this manual before using j the rotor 1 Inspect the O rings and plunger mechanism for damage the high forces generated in this rotor can cause damaged components to fail 2 Use only tubes and accessories listed in Table 1 3 Check the chemical compatibilities of all materials used refer to Appendix A in Rotors and Tubes TLA 120 2 Rotor ROTOR PREPARATION OPERATION Plunger Engaged uum Tm
30. trifuges The rotor is made of titanium and is finished with black polyurethane paint The lid is made of aluminum and anodized to resist corrosion A plunger in the lid locks the rotor to the drive hub before beginning the run and two lubricated O rings made of Buna N rubber maintain atmospheric pressure inside the rotor during centrifugation The tube cavities are numbered to aid in sample identification This rotor was tested to demonstrate containment of microbiological aerosols under normal operating conditions of the associated Beckman Coulter centrifuge when used and maintained as instructed The rotor is specially designed with a fluid containment annulus located below the O ring sealing surface see Figure 1 The annulus retains fluid that may escape from leaking or overfilled tubes thereby preventing the liquid from escaping into the instrument chamber O ring Sealing Surface Annulus Figure 1 Fluid Containment Annulus Validation of microbiological containment was done at an independent third party testing facility CAMR Porton Down UK or USAMRIID Ft Detrick MD U S A Improper use or maintenance may affect seal integrity and thus containment TLA 120 2 Rotor The ultracentrifuge identifies rotor speed during the run by means of a magnetic speed sensor in the instrument chamber and magnets on the bottom of the rotor This overspeed protection system ensures that the rotor does no
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