User Manual - Galileo Bioscience
Contents
1. Water distilled JJ 73 NN 22 22 22 22 22 22 22 22 22 22 22 22 22 72 22 22 22 22 22 272 22 22 72 22 22 22 22 22 22 22 22 22 22 232 Chromic Acid over 4099 Chloroform Diethyl Ether Dimethyl Formamide Ethyl Acetate Ethyl Alcohol 9599 Ethylene Dichloride Hydrofluoric Acld 4099 Hydrogen peroxide over 4099 Lacquer Thinner Methyl Alcohol over 1599 Methylene Chloride Methyl Ethyl Ketone MEK Nitric Acid concentrate Phenol 5 solution Sulfuric Acid concentrate Toluene Trichloroethylene Xylene Copyright Galileo Bioscience 2004 Page 18 NOTES Copyright Galileo Bioscience 2004 Page 192. Problem Bands are Narrower than the Sample Wells gt lonic strength of the sample is higher than that of the gel De salt the sample or use sample buffer of the same strength as the gel Problem Distorted Sample Wells gt Incomplete polymerization produces poorly defined wells De gas gel solution prior to casting and increase APS and TEMED concentrations The comb can be wiped with TEMED just prior to casting to improve polymerization gt Salt concentration is too high in the sample Dialyze sample or use desalting column Problem Resolving gel is uneven at the top gt Overlay gel carefully using water saturated n butanol and make sure casting stand is level Problem Poorly Resolved Bands May be caused by too much sample for well width or gel thickness Dilute sample Lower volumes generally give better resolution gt Excessively high voltages cause fast run times but poor resolution Sample may have degraded Problem Frowning of Outside Lanes gt Leakage of Buffer along the sides or along the spacers inside the gel assembly Do not move spacers after polymerization and make sure that the gasket is seated firmly against the glass Problem Double Bands Doublets gt Due to re oxidation or insufficient reduction of the sample If using a reducing agent prepare fresh sample buffer every 30 days Increase the concentration of 2 mercaptoethanol or dithiothreitol in the sample Problem Fewer Bands than Expec
3. Resolve to Discover the syzygy of science art amp practicality GALILEO bioscience vt ia i r RAE RR AR A 7 WA Um mE d dl P p TR a i C BA s p MANUAL 1 47S O 1 an rr ff E B N 4 it A AJ A WP V S A ON b Ww Table of Contents COVE PAGO r EU DIU TM CP EOO E E E ee eee TOC Tithe Page Contact INO wicceeistcevesacctcgeeteseeweieeweteaceeie von ei 1 Warranty Component IIlUStration ccc ccc eee cece cease eases eaeeeesaaees 2 m ieXnoBLicm an 3 4 Environmental Conditions Safety Precautions 5 Instructions For Use NWT OUST ON ARENA RD NONE IDR TORRE 6 odieb PIeCANHONS css serao edm Far E E Wm panelas dei MM HEUS 7 General Specifications amp Operating Parameters 7 General Care amp Cleaning cece ccc eee ee ee eee eeesseeeeseesseessaeeees 8 Gel Plate Preparation cccccc ccc cece ence eee sense eee eeeaeeeeeaseensaeneees 8 Gel Plate Cassette ASSEMDIY 0 0 cece cece seen 8 Unit Assembly For Casting and Running Gels 9 Casting WG OO PDR O 10 11 Gel and Buffer VolUUSuesscdedder n dre pe scans aceon EATEN eai 11 Sample Loading cece cece ee ee cece eens eee esas ee eeeeeaeeeseeaaee
4. power cords to the power supply STARTING ENDING THE RUN STARTING ENDING Connect the chamber to the power supply and connect the power supply to the main electrical source Turn all settings to zero before turning on the main source of electricity Adjust the controls to the desired settings Follow manufacturer s instructions Turn power supply settings to zero turn off the main electrical source and disconnect the power cords Turn off the water if using optional cooling Remove the lid by pushing on the acrylic alignment pins protruding through the top of the lid with your thumbs Slide and lift the upper buffer chamber out of the lower buffer chamber and drain buffer chambers separately Copyright Galileo Bioscience 2004 Page 13 gt Loosen wing nuts and slide clamp bars outward to remove gel cassettes It is not necessary to remove the clamp bars from the upper buffer chamber to remove the gel cassette gt After the gel cassette s has been removed the gel s are ready for staining and blotting Separate the plates with a strong broad blade When using notched glass plates DO NOT pry them apart at the notches Spread the load over a wide area gt Rinse the chambers with distilled water then dry the electrode connectors with tissue Ensure that the connectors are clean and dry before usage or storage TROUBLESHOOTING Many factors may affect the quality of vertical gel preparations For example prepar
5. mixture Copyright Galileo Bioscience 2004 Page 16 10 0 5 0 10 0 5 0 6 0 6 0 0 3 0 3 12 2 17 2 1 5 1 5 0 015 0 015 REPLACEMENT PARTS DESCRIPTION Power Supply Cords Retractable Sheath Power Supply Cords Non Retractable Sheath UBC Replacements Gaskets Casting Base Replacement Gaskets Blank Glass Plate 3 32 2 3mm Blank Glass Plate 1 8 3 0mm Notched Glass Plate 3 32 2 3mm Notched Glass Plate 1 8 3 0mm Notched Alumina Plate 1 0mm Blocking Plate for Running One Gel Spacers 0 8mm Spacers 1 5mm Clamp Assemblies Wing Knobs 4 Spring Pegs 2 Positioning Pegs Positioning Peg Gasket 85 1010 RPC NRPC 85 1010 GSK 85 1010 CBGK 85 1010 10BG 85 1010 10NG 85 1010 10NA 85 1010 BPL 85 1010 SP 0 8 85 1010 SP 1 5 85 1010 CL 85 WKNB 85 PEGS 85 PP 85 PP GK 85 2010 RPC NRPC 85 2010 GSK 85 2010 CBGK 85 2010 10BG 85 2010 10NG 85 2010 BPL 85 2010 SP 0 8 85 2010 SP 1 5 85 2010 CL 85 WKNB 85 PEGS 85 PP 85 PP GK Copyright Galileo Bioscience 2004 Page 17 85 1614 RPC NRPC 85 1614 GSK 85 1614 CBGK 85 1614 14BG 85 1614 14NG 85 1614 BPL 85 1614 SP 0 8 85 1614 SP 1 5 85 1614 CL 85 WKNB 85 PEGS 85 PP 85 PP GK 85 2020 RPC NRPC 85 2020 GSK 85 2020 CBGK 85 2020 20BG 85 2020 20NG 85 2020 BPL 85 2020 SP 0 8 85 2020 SP 1 5 85 2020 CL 85 WKNB 85 PEGS 85 PP 85 PP GK The following is a chemical compatibility chart for the care o
6. should rest on the positioning squares of the Casting Base and between the nylon alignment pins 5 To clamp the assembly to the Upper Buffer Chamber slide the clamp bars towards the middle Tighten the wing nuts until a seal is formed between the gasket and the glass The hollow gasket allows for a superb seal without over tightening the wing nuts Over tightening may cause the glass to break 6 If casting and running two gels repeat steps 4 6 for the second gel plate assembly If you desire to run only one gel secure the blocking plate to the second side Note a combination of two gel assemblies or one gel assembly and the Blocking Plate are necessary to form the walls of the Upper Buffer Chamber 7 Lift the assembled Upper Buffer Chamber and turn the Casting Base over Turn the cams so that the handle is pointing up and pull out Place the Upper Buffer Chamber on the gaskets Note A protective plastic film is left on the gaskets for shipping A piece of clear tape has been fastened to the end of the film to assist the user in removing it 8 Insert the cam pins and simultaneously turn the handles one half turn to tighten the assembly down onto the gasket base Once the Upper Buffer Chamber assembly has been secured onto the casting base an initial leak test using a small amount of water is recommended add 2ml 3ml of water and let stand for 2 minutes If no leakage is visible empty water and proceed with gel casting Copyrig
7. the gel assembly is seated firmly against the gasket Remove gasket wash in warm water to remove excess salts and place the gasket back in the groove Running at too low a current Use running conditions given in this manual When running at constant current the current value listed is per gel Problem Running Too Fast gt Check buffer recipe remake and try again If voltage is lower than usual when running at constant current the buffer is probably too dilute Copyright Galileo Bioscience 2004 Page 14 gt Voltage or current may be set too high turn down current setting Problem Smiling of Dye Front gt The center of the gel is running hotter than at the edges use coolant or cold tap water in cooling core and or turn down current setting Problem Vertical Streaking gt Excessive sample or particles in the sample either dilute sample or reduce voltage Centrifuge Samples to remove particulate contamination gt Sample has precipitated Centrifuge sample before adding sample buffer or use a lower acrylamide gel Problem Bands Spread Laterally gt Diffusion of sample Make sure the samples are loaded quickly and the power Is applied as soon as possible after loading gt Diffusion of sample during the run in the stacking gel Increase of stacking gel or increase current by 25 when stacking gt Lower ionic strength of the sample Match the ionic strength of the sample with that of the gel
8. 2004 Page 6 USING THE VERTICAL GEL ELECTROPHORESIS UNITS gt D re amp Pe SAFETY PRECAUTIONS READ all instructions before using the unit Always turn off power supply FIRST then disconnect the power cords Always have electrophoresis unit disconnected from their power supply before removing the safety cover DO NOT exceed the maximum operating voltage or current see TABLE A DO NOT operate electrophoresis units in metal trays Acrylamide is a volatile cumulative neurotoxin and suspected carcinogen Wear effective protective clothing and follow recommended handling and disposal procedures Polymerized gels contain some unpolymerized monomer Handle with gloves only DO NOT fill the unit with running buffer above the maximum Fill Line DO NOT move the unit when it is running CAUTION During electrophoresis very low quantities of various gases are produced at the electrodes The type of gas produced depends on the composition of the buffer employed To disperse these gases make sure that the unit is run in a well ventilated area TABLE A General Device Specifications General Operating Parameters 85 1010 NCS 85 2010 NCS Unit Model Number 85 1010 WCS 85 2010 WCS 85 1614 85 2020 10cmW x 10cmL 16cmW x 14cmL Gel Size Or 20cmW x 10cmL Or 20cmW x 20cmL 10cmW x 8cmL 16cmW x 16cmL Upper Buffer Chamber 170ml non cooled 400ml non cooled 4 Capacity slightly more slightly more 400ml 600ml pay mom as0m 300m Capac
9. Width Catalog of Thickness of Teeth EST Well Number Teeth of Tooth mm mm Volume ul 85 2020 C10 0 8 10 0 8 13 6 239 85 2020 C10 1 5 85 2020 c15 0 8 15 08 82 M 85 2020 c20 0 8 20 08 55 9 0 1 5 2 9 182 85 2020 C20 1 5 85 2020 C25 0 8 85 2020 C25 1 5 85 2020 CPREP COOLING OPTION As previously noted all four sizes of Galileo Reflection Twin Vertical Electrophoresis Units are available with optional water cooling Sometimes cooling is needed when running gels at higher current or when the bioactivity of an enzyme has to be preserved Heating of the gel can cause smiling and other problems with the resolution of protein bands This is particularly pronounced on larger gels We recommend running coolant or water through the cooling core in the upper buffer chamber When ramping up voltage or current consider at least tap water cooling TO UTILIZE OPTIONAL COOLING Attach a separate piece of 3 8 ID clear flexible lab tubing to each hose barb on the upper buffer chamber marked as in and out Attach the tubing from the cathode black side of the unit marked in to either a cold water tap or a re circulator chiller Water flow should not exceed 2 L per minute at 30 psi Attach the drain tube on the anode red side marked as out to the re circulator chiller or put into the sink drain Turn on the water Once the water has started to circulate through the system connect the
10. ation of gel and sample buffers gel casting and tank assembly and or run conditions Reading and following the instructions in this operating manual can solve most problems Below we list some of those most commonly experienced problems along with suggestions for solving them Problem Acrylamide Solution Leaks During Casting gt Ensure that the sealing surfaces of the glass plates and spacers are clean gt Ensure that each plate is free of chips gt Ensure that the wing knobs on the upper buffer chamber are tightened use care not to over tighten gt Ensure that the glass plates and spacers have been set in place using the positioning squares on the Flip Side of the Gel Casting Base gt Ensure that the Cams in the Casting Base have been turned equally to tighten down the Upper Buffer Chamber on the gaskets Problem Bubbles Do Not Appear on the Electrodes gt Check to see if the Power Supply is operating properly Problem Gels Fail to Polymerize gt May be caused by low temperatures oxygen insufficient degraded catalyst or low acrylamide concentrations Problem Run Takes Longer Than Usual gt Buffers may be too concentrated or at the wrong pH Gel concentration may be too high Check Buffer Recipe and try again See if voltage produced by the current you are running at is the same lf it differs significantly your buffer may not have been made up correctly Upper Buffer Chamber may be leaking buffer Make sure
11. e operated safely at an altitude of 2 000 m The normal operating temperature range is between 49C and 659C gt Maximum relative humidity 80 for temperatures up to 319C decreasing linearly to 50 relative humidity at 409C SAFETY PRECAUTIONS Please read the User Manual carefully before using the Vertical Electrophoresis Unit This manual contains important operating and safety information Our electrophoresis units are designed to perform flawlessly for years in the most demanding laboratories Please take the time to read the manual to ensure that you understand the safety and operating instructions to ensure the successful use of the unit Alterations could cause serious injury to the user or the system Power to the unit is supplied by an external power supply The power supply must meet safety standards for IEC 1010 1 regulations and must be ground isolated and incorporate a no load detecting circuit Power is supplied to the gel through the lid of the system providing a safety interlock to the user Users should not attempt to operate this unit without the safety inter locked lid in place A Always disconnect the unit from the power supply before removing the cover to avoid the risk of personal shock A Running Conditions should not exceed the maximum operating voltage or current A Do not fill the Lower Buffer Chamber with running buffer above the maximum fill line A Always disconnect the unit from the power supply when you want to mo
12. f acrylic Acrylic is compatible with most solvents and solutions found in a biochemical laboratory but some solvents can cause damage The list does not include all possible chemical compatibilities and or safe compounds R RESISTANT LR LIMITED RESISTANCE N NOT RESISTANT LISTED BY CODE ALL CHEMICALS IN THE FIRST COLUMN ARE SAFE TO USE Chemical Code Chemical Code RESISTANT LIMITED RESISTANCE Acetic Acid 5 Ammonia Acetic Anhydride LR Ammonium Chloride saturated Chromic Acid up to 40 LR Ammonium Hydroxide 10 Dioctyl Phthalate LR Ammonium Hydroxide concentrate Ethyl Alcohol up to 30 LR Battery Acid Gasoline regular leaded LR Calcium Chloride saturated Isopropyl Alcohol up to 5099 LR Calcium Hypochlorite Methyl Alcohol up to 1599 LR Citric Acid 2099 Nitric Acid 20 7099 LR Cottonseed Oil edible Detergent Solution Heavy Duty Diesel Oil NOT RESISTANT Acetic Acid Glacial 2 Ethylhexyl Sebacate Acetone Ethylene Glycol Aniline Formaldehyde 4099 Benzene Glycerine Butyl Acetate Heptane Carbon Tetrachloride Hexane commercial Grade Hydrochloric Acid 1099 Hydrochloric Acid concentrate Hydrogen peroxide up to 4099 Hydroxide 1099 Isooctane Kerosene no 2 fuel oil Mineral Oil white Naphtha VM amp P Nitric Acid up to 2099 Oleic Acid Olive Oil Soap Solution lvory Sodium Carbonate Sodium Chloride Sodium Hydroxide Sodium Hypochlorite Sulfuric Acid up tp 3099 Turpentine
13. f the Upper Buffer Chamber that slide into the precision machined clear sides of the Lower Buffer Chamber to set it in place Copyright Galileo Bioscience 2004 Page 10 4 Add the appropriate volume of running buffer to the upper buffer chamber Table A gives approximate volumes making sure the running buffer is 3 mm below the top of the blank glass ensuring sufficient contact with the top of the gel surface Be sure that the running buffer is not leaking from the upper buffer chamber to the lower buffer chamber If buffer is leaking you will need to drain the UBC and reset the gel cassettes NOTE When running only one gel a blocking plate is required on the other side of the unit to retain the top buffer level GEL AND BUFFER VOLUMES Some guidelines for general operating conditions are given in Table A but conditions vary according to the number of gels their composition length and cross sectional area The current requirement will increase in proportion to the number of gels or gel thickness providing that the voltage is not limiting e g 2 gels require twice the current of 1 but the same voltage Longer gels require proportionally higher voltages By increasing the gel concentration the electrical resistance is increased and the rate of migration decreases Higher voltages can be applied but be careful not to overheat the gel The conductivity of non dissociating buffer systems gels vary enormously and conditions must be determined em
14. ge 11 TABLES C1 C2 C3 amp C4 Comb Options Table C1 MODEL 85 1010 NCS or 85 1010 WCS Combs Number Width ak o E a EST ay of Tooth Volume mme os fae 8 100615 6 15 Hl esamoceos e os n 9 85 1010 c8 1 5 8 15 77 185 851010MT9 8 9 08 66 8 85 1010 CMT9 1 5 9 15 66 160 85 1010 c10 0 8 10 08 57 7 85100 1208 12 08 43 ss 8100 1215 12 is 43 o mmmocisos s os 30 f s Table C2 MODEL 85 1614 Combs ENT Width Catalog di of Teeth EST Well TCU THES sth of Tooth a7 mm Volume ul 85 1614 C10 8 183 85 1614 C10 1 5 CETTE TO GR O a 85 1614 C15 0 8 5 os 1 17 EPH l6lAV CIS8 15 15 61 20 EPH 1614v c20 08 20 os 39 6 amp 6 EPH I614V C4 08 24 08 29 5 EWieMvOMis 26 18 Las __ _96__ ps 1094 47 3630 152 Table C3 MODEL 85 2010 NCS or 85 2010 WCS Combs Width Catalog Number EE 2 d EST Mor Number of Teeth of Tooth Volume Tal a LM a 85 2010 C10 1 5 10 15 136 449 85 2010 C15 0 8 15 os 82 qua 8 200 155 15 15 82 2n 852010 CMT18 8 18 os 65 78 85 2010 CMT18 1 5 SE 85 2010 C20 0 8 85 2010 C25 0 8 85 2010 C25 1 5 as 39 x 85 2010 CMT36 0 8 mas 08 21 85 2010 CMT36 1 5 Copyright Galileo Bioscience 2004 Page 12 Table C4 MODEL 85 2020 Combs Number
15. ht Galileo Bioscience 2004 Page 9 Upper Buffer Chamber Fr Reversed l Casting Base Mf Upper Buffer Chamber Alignment Pins em nus a x9 Positioning Squares Casting Base E Blocking Plate Plate Assembly Upper Buffer Chamber gt P e pi Casting Base Sd NE gt p CASTING THE GEL To ensure reproducibility and uniform polyacrylamide cross linking we recommend de ionizing de gassing and filtration of acrylamide gel solutions prior to use Acrylamide solutions should be stored in a cool dark environment such as a refrigerator and allowed to reach room temperature prior to pouring Avoid exposure to heat and sunlight Polymerization conditions should be adjusted to effect polymerization within about 5 10 minutes Test a small volume in a vial prior to pouring the gel As a rough guide 100ml of degassed 6 acrylamide gel will set in about 5 minutes at room temperature when gently mixed with 450ul of freshly prepared 10 w v Ammonium persulphate plus 200ul TEMED The setting time increases to about 10 minutes if the TEMED volume is reduced to 100ul and to approximately 15 minutes with 75ul The amount of catalysts may need to be reduced under warm conditions Do not pour under direct sunlight 1 Prepare the appropriate volume of acrylamide gel solution using Table B below as a guide These volumes have been calculated using the glass and spacers provided by Galileo a
16. ick Notched Alumina Plates 10cmW x 10cmL x 0 10cm thick Blocking Plate for running one gel Side Spacers 0 8mm thick 10 well Teflon amp Comb 0 8mm thick Set 2 replacement gaskets for the upper buffer chamber Casting Base Set 2 replacement gaskets for the casting base 85 2010 WCS amp 85 2010 NCS 1 1 1 1 4 4 1 8 2 2 1 1 1 85 1614 User s Manual Safety Lid with integral double insulated power cords rated safe up to 1 000 volts with Retractable Sheathed Power Connectors Upper Buffer Chamber with color coded electrodes gold plated banana plugs 85 2010 WCS units has a cooling core sealed with an alumina plate amp in and out Hose Barb Fixtures for water circulation Lower Buffer Chamber Blank Glass Plates 20cmW x 10cmL x 0 32cm thick Notched Glass Plates 20cmW x 10cmL x 0 32cm thick Blocking Plate for running one gel Side Spacers 0 8mm thick 15 well Teflon amp Comb 0 8mm thick 20 well Teflon Comb 0 8mm thick Set 2 replacement gaskets for the upper buffer chamber Casting Base Set 2 replacement gaskets for the casting base Copyright Galileo Bioscience 2004 Page 3 1 1 1 1 4 4 1 8 2 2 1 1 1 85 2020 1 1 1 1 4 4 1 8 2 2 1 1 1 User s Manual Safety Lid with integral double insulated power cords rated safe up to 1 000 volts with Retractable Sheathed Power Connecto
17. ity 240ml 450ml 300ml 800ml Total Running Buffer 450ml 750 ml 650ml 1250ml Total Buffer Capacity 450ml 1100 1300 ml 650ml 1250ml Current mAmps Constant 15 35mA gel 30 45mA gel 15 50mA gel 15 75mA gel Maximum Voltage volts 600V 600V 600V 600V Time Requirements 30 90 minutes 30 90 minutes 60 120 minutes 60 180 minutes Sample Capacity 24 72 48 20 Copyright Galileo Bioscience 2004 Page 7 GENERAL CARE amp CLEANING WARNING Acrylic is not resistant to aromatic or halogenated hydrocarbons ketones or esters Organic solvents cause acrylic to craze or crack Do not use ethanol or other organic solvents to clean your unit Do not autoclave bake or microwave your unit A Before using clean and dry unit with DISTILLED WATER ONLY dry parts with clean tissues or air dry Use care when cleaning or drying the unit near the platinum wire The connectors should be clean and dry before usage or storage Do not use abrasive creams or scourers A Do not use cleaning brushes in the electrode area A Athorough rinse with distilled water is all that is generally required to clean the unit after use A mild detergent may also be used Acrylic can also be exposed to a mild bleach solution 10 1 In addition RNAse removal products are also safe for acrylic See Page 19 for Chemical Compatibility Chart GEL PLATE PREPARATION Clean the glass plates spacers and combs in mild labo
18. lignment Pins amp Hollow Gaskets Guarantee Reliable Leak Proof Gel Installation gt Precision Glass Plates Provide Exceptional Flatness and Finished Edges to Ensure Uniform Separation gt Casting Base Enables Casting Directly on the Upper Buffer Chamber Obviating Need to Move Gels once Polymerized Intelligent Design Results in Exceptional Resolution gt Electrode Configuration Assures Uniform Field Straight Lanes and Rapid Runs Saving Time and Improving Data Generation Rate gt Proximal Upper Buffer Chamber Exploits Specific Heat of Aqueous Buffer to Provide Uniform Temperature and No Smiling gt Efficient Water Cooling System Available on All Systems Prevent Band Distortion gt Optional Notched Alumina Plates available for the mini 10 cm x 10cm unit Enhance Heat Dissipation Wide Variety of Options Maximize Product Versatility gt Devices Available For Four Gel Sizes Including the wide mini 85 2010 WCS amp 85 2010 WCS that Accommodate 72 Samples Supporting Most PAGE Applications gt Optional Additional Upper Buffer Chambers Allow for Simultaneous Use of the Twin Systems Improving Data Output Rate gt Non Cooled 10cm x 10cm amp 20cm x 10cm Upper Buffer Chambers Available gt Wide Selection of Combs plus Glass and Blocking Plates Available for All Units gt Units are Compatible with Pre cast Acrylamide Gels from Most Manufacturers and Vertical Agarose VAGE Separation Copyright Galileo Bioscience
19. lileo Bioscience Galileo guarantees that the Vertical Electrophoresis System you have received has passed rigorous quality assurance protocols and meets its published specification This warranty is valid for 48 months only if the product has been used and cared for according to this user manual No liability is accepted for loss or damage arising from incorrect use Galileo s liability is limited to the repair or replacement of the unit or refund of the purchase price at our option Galileo is not liable for any consequential damages Galileo reserves the right to alter the specifications of the Vertical Electrophoresis Systems without prior notice This will enable us to implement improvements as soon as they become available VERTICAL ELECTROPHORESIS SYSTEM COMPONENTS To Power Supply EN Casting Base Copyright Galileo Bioscience 2004 Page 2 PACKING LISTS 85 1010 WCS amp 85 1010 NCS 1 1 1 1 4 2 2 1 4 2 1 1 1 User s Manual Safety Lid with integral double insulated power cords rated safe up to 1 000 volts with Retractable Sheathed Power Connectors Upper Buffer Chamber with color coded electrodes and gold plated banana plugs 85 1010 WCS unit has a cooling core sealed with an alumina plate amp in and out Hose Barb Fixtures for water circulation Lower Buffer Chamber Blank Glass Plates 10cmW x 10cmL x 0 24cm thick Notched Glass Plates 10cmW x 10cmL x 0 24cm th
20. nd subtracting the volume of the spacers and the notch The volumes are approximate TABLE B Approximate Gel Solution Volumes For Various Cassette Configurations Unit Plate Width Plate Length Spacer Thickness Gel Volume 85 1010 10cm 10cm 0 8mm 7 5m 10cm 10cm 1 5mm 15ml 10cm acm 0 8mm 6ml 10cm 8cm 1 5mm 12ml 85 2010 20cm 10cm 0 8mm 15ml 20cm 10cm 1 5mm 30ml 85 1614 16cm 14cm 0 8mm 13 5ml 16cm 14cm 1 5mm 27ml 16cm 16cm 0 8mm 15ml 16cm 16cm 1 5mm 30 ml 85 2020 20cm 20cm 0 8mm 24 6ml 20cm 20cm 1 5mm 49 1ml 2 Run the acrylamide gel solution mix slowly down the inside edge of the gel cassette Avoid aeration Place the comb in the gel plate assembly a If a stacking gel is to be used carefully overlay the gel solution to a depth of 3 5mm with 1x gel buffer or water saturated butanol Following polymerization of the separating gel pour off the overlay layer rinse off butanol with electrophoresis gel buffer and pour a stacking gel if required Insert the comb ensuring bubbles are not trapped around comb teeth Once the stacking gel has polymerized use the gel immediately or store wrapped in a damp paper towel and plastic film at 4 C9 Wait a minimum of 15 minutes for the gel to polymerize Repeat process as required 3 Release the cams and pull away from the Upper Buffer Chamber and gels Wash off any residual acrylamide Place the Upper Buffer Chamber into the Lower Buffer Chamber Stainless Steel pins are located on the lower sides o
21. pirically The run conditions are to be taken as a guideline only and apply to SDS Tris glycine gels If the plates become hot increase the water flow rates within the recommended limits or reduce the power settings SAMPLE LOADING gt If a native gel is being used pre electrophorese the gel for 15 40 minutes prior to loading samples SDS gels do not need this step gt Centrifuge samples at 12 000 x g for 5 minutes If this step is omitted samples may streak during electrophoresis gt Carefully remove the sample comb and immediately flush the wells with electrophoresis buffer using a syringe Load the samples using a gel loading pipette tip See TABLE D on the next page for approximate well volumes etc If possible avoid taking liquid from the pellet area at the bottom of the tube During sample loading the pipette tip should be 1 2 mm above the bottom of the well to minimize dilution of the sample and to keep the sample as a tight layer gt Fill unused wells with the equivalent volume of sample buffer to maintain uniform electrical resistance across the gel gt Add buffer to the lower buffer chamber to approximately 2 3 mm above the base of the gel using the Fill Line as a guide The bottom end of the gel assembly should be in contact with the running buffer Set the safety lid onto the unit so that the power cords are connected in the proper position red to red black to black Copyright Galileo Bioscience 2004 Pa
22. ratory detergent A DO NOT use abrasive creams or scourers If a particularly clean finish is required e g for silver stained gels glass plates can be soaked in chromic acid overnight rinsed with water then wiped successively with ethanol acetone then ethanol again A DO NOT ALLOW organic solvents or chromic acid to come into contact with the acrylic components of your vertical system Handle clean plates with gloved hands remove any finger prints with acetone GEL PLATE CASSETTE ASSEMBLY 1 Ona clean level bench position the two side spacers flush with the edges of the blank glass plate and overlay the notched plate Copyright Galileo Bioscience 2004 Page 8 UNIT ASSEMBLY FOR CASTING AND RUNNING GELS 1 Turn Over the Casting Base so that the four acrylic positioning squares are on the top 2 Place the Upper Buffer Chamber on the Casting Base The precision machined Upper Buffer Chamber should fit over the four squares snugly 3 Loosen the wing nuts and slide the clamp bars outward Please note the nylon alignment pins These pins assure that the glass plates are properly placed over the Upper Buffer Chamber gasket while the acrylic squares on the casting base assure that the glass is positioned evenly and at the proper place for optimum sealing on the casting base gaskets 4 Place the gel plate assemblies cassettes with the notched glass plate innermost against the gasket of the Upper Buffer Chamber The assemblies
23. rs Upper Buffer Chamber with color coded electrodes and gold plated banana plugs 85 1614 units have a cooling core sealed with an alumina plate amp in and out Hose Barb Fixtures for water circulation Lower Buffer Chamber Blank Glass Plates 16cmW x 14cmL x 0 32cm thick Notched Glass Plates 16cmW x 14cmL x 0 32cm thick Blocking Plate for running one gel Side Spacers 1 5mm thick 15 well Teflon amp Comb 1 5mm thick 20 well Teflon amp Comb 1 5mm thick Set 2 replacement gaskets for the upper buffer chamber Casting Base Set 2 replacement gaskets for the casting base User s Manual Safety Lid with integral double insulated power cords rated safe up to 1 000 volts with Retractable Sheathed Power Connectors Upper Buffer Chamber with color coded electrodes and gold plated banana plugs 85 2020 units have a cooling core sealed with an alumina plate amp in and out Hose Barb Fixtures for water circulation Lower Buffer Chamber Blank Glass Plates 20cmW x 20cmL x 0 32cm thick Notched Glass Plates 20cmW x 20cmL x 032cm thick Blocking Plate for running one gel Side Spacers 1 5mm thick 15 well Teflon Comb 1 5mm thick 20 well Teflon Comb 1 5mm thick Set 2 replacement gaskets for the upper buffer chamber Casting Base Set 2 replacement gaskets for the casting base Copyright Galileo Bioscience 2004 Page 4 ENVIRONMENTAL CONDITIONS FOR USE gt This unit is intended for indoor use only gt This unit can b
24. ssneeees 11 eodein M QU 12 13 OO III CNG n PER e ENO RR 13 Starting Ending The Run eee erre 13 14 TOUDI ONOONO ssa RETTULIT T 14 15 Reagent Information ccc errar rear 16 Repiacement PAN asneiras ao eia dar a ita 17 Chemical Compatibility Chart cc serrana 18 NOTES us ssa se ss E E A EE 19 Copyright Galileo Bioscience 2004 TOC TWIN VERTICAL ELECTROPHESIS GALILEO o o USER S MANUAL rev 10 1 04 bioscience 85 1010 WCS 85 1010 NCS 85 2010 WCS 85 2010 NCS 85 1614 AND 85 2020 A mme A THESE UNITS ARE CAPABLE OF DELIVERING POTENTIALLY LETHAL VOLTAGE WHEN CONNECTED TO A POWER SUPPLY AND ARE TO BE OPERATED ONLY BY QUALIFIED TECHNICALLY TRAINED PERSONNEL PLEASE READ THE ENTIRE OPERATOR S MANUAL THOROUGHLY BEFORE OPERATING THIS UNIT UNITS COMPLY WITH THE STATUTORY CE SAFETY DIRECTIVES Galileo Bioscience P O Box 390566 Cambridge MA 02139 Toll Free 877 481 9175 Tel 781 481 9175 Fax 781 481 9214 www galileobioscience com Copyright Galileo Bioscience 2004 Page 1 g WARRANTY Please check that the unit has been received complete and undamaged Refer to the Packing Lists on pages 3 amp 4 and check that all components and accessories are present Be sure to save all packaging and documents until you have thoroughly inspected your shipment if you should find that your order is incorrect or damaged call for return instructions Ga
25. ted with Heavy Band at Dye Front gt Caused by more than one band migrating to the dye front increase total monomer concentration Yol gt Sample may have degraded due to incorrect storage and or contamination Copyright Galileo Bioscience 2004 Page 15 REAGENT INFORMATION RUNNING BUFFER TGS Tris 3 02859 L Glycine 14 4g L SDS 1 0g L pH 8 3 Laemmli 1970 Q s to 1L Note For Native Protein Electrophoresis do not add SDS Table D1 Sample Buffer 2X Concentration Final Concentration Stock With Sample L 10mL 2 SDS 20g 0 2 1 10 BME lOomL 0 1 5 25mM Tris 6 057g 0 0606g 125mM 30 Glycerol 300mL 3mL 15 0 002 Bromo Phenol Blue 02g 0002g 0 001 add sample buffer 1 1 with sample solution Caution 2X Sample Buffer containing 2 mercaptoethanol should be prepared in a fume hood 0 2M final concentration Dithiothreitol DTT may be used in place of 2 mercaptoethanol DDT should be added before use and made fresh ACRYLAMIDE SOLUTION Stock acrylamide solution for D2 29 2g Acrylamide and 8 bis Acrylamide q s 100mL H20 TABLE D2 Gel Preparation SDS Page continuous buffer system 9o Acrylamide Stock Solution 20 0 15 0 12 5 Acrylamide Bisacrylamide 30 08 20 0 15 0 12 5 0 5 M Sodium Phosphate Buffer pH 7 2 6 0 6 0 6 0 10 w v SDS 0 3 0 3 0 3 Water 2 2 12 9 7 1 596 w v APS 1 5 1 5 1 5 TEMED 0 015 0 015 0 015 The columns represent volumes ml of stock solutions required to prepare 30ml of gel
26. ve the unit or add running buffer A Use this apparatus only for its intended purpose as described in this manual Do not use this product if the power cords are damaged or if any of its surfaces are cracked Copyright Galileo Bioscience 2004 Page 5 Reflection Twin Vertical GALILEO Series INSTRUCTIONS FOR USE Q INTRODUCTION Thank you for your purchase of a Galileo Reflection Twin Vertical Gel Electrophoresis System Our vertical systems allow for fine resolution of protein or nucleic acid fragments on one or two acrylamide gels PAGE PAGE separation offers the superior resolution necessary to separate native or denatured proteins and nucleic acids in applications such as SSCP or dinucleotide repeat analysis using western blotting and also for automated protein sequencing analysis All four models in the Galileo Reflection Twin Vertical Series incorporate inspired design features and exceptional manufacturing methods that ensure dependable performance over years of continuous use A comprehensive offering of combs and accessories plus the compatibility of the 85 1010 WCS amp 85 1010 NCS with most commercially available pre cast mini gels ensures maximum system utility to exceed the separation demands of most research laboratories Outstanding Features Ensure Trouble Free Use gt Robust Acrylic Construction Stands up to Daily Usage without Breakage Warping or Leakage gt Rugged Spring Loaded Clamp Mechanism A
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