Eco-Line user manual (English)
Contents
1. Acidic buffer system transfer towards cathode Place transfer membranes on cathode side lid and gel on anode side base Incomplete transfer Transfer time too short check by gel staining Increase transfer time Transfer time too long blow through i e proteins are stained on both sides of the membrane Reduce transfer time Use nitrocellulose with smaller pore size or PVDF membrane Irregular transfer Air bubbles in the blot sandwich Remove air bubbles by gently rolling e g a pasteur pipette over the membrane Generation of air bubbles with long transfer times by electrolysis and heating up Reduce transfer time chill buffer before use Irregular current flow as size of blotting papers and transfer membrane are larger than gel size Transfer membranes and blotting papers must be cut exactly to gel size Blotting paper partially dried Reduce transfer time Insufficiently soaked transfer membrane Check procedure for prewetting membrane Spoiled electrodes Clean electrodes Proteins are stained on both sides of the membrane Transfer time to long blow through Reduce transfer time Use nitrocellulose with smaller pore size or PVDF membrane Power Supply turns off Power Supply does not work at low voltages inner resistance too high Use proper Power Supply which can operate at low voltages e g Standard Power Pack P25 Stro2. Warning The apparatus must not be earthed Eco Line Manual Vers 2014 06 16 4 Components 4 1 Systems Overview Eco Mini System E EB or EBC Eco Maxi System EB or EBC Buffer Tank E EB or EBC e Bigfoot Safety Lid with power cords e Electrophoresis Module e Casting stand Eco Line Manual 2014 06 analytikjena Mo Picture Buffer Tanks Eco Mini Picture Bigfoot Safety Lid Eco Mini Picture Casting Stand Eco Mini left and Eco Maxi right Biometra analytikjena Tankblot Eco Mini or Eco Mini C Tankblot Eco Maxi or Eco Maxi C Buffer Tank EB or EBC e Bigfoot safety lid with power cords e Blot Module e Blotting Cassettes colour coded 17 ibs Picture Buffer Tanks Eco Mini Picture Bigfoot Safety Lid Eco Mini Picture Blot Module Eco Mini left and Eco Maxi right Picture Blotting Cassette Eco Mini left and Eco Maxi right Eco Line Manual Vers 2014 06 18 analytikjena 4 2 Compatibility Overview Buffer tank E for electrophoresis Eco Mini only Buffer tank EB for electrophoresis and blotting Buffer tank EBC for electrophoresis and blotting with integrated cooling Siometr Eco Line Manual 201406 analytikjena m 5 Electrophoresis 5 1 Gel Cassette Assembly A Clean and dry glass plates before use e Single gel sandwiches Select a spacer plate of the desired gel thickness and place the notched glass plate o
3. Current of voltage set too high Turn down electrical settings Smiling of the dye front Center of the gel is hotter than the sides Turn down electrical settings Biometra Ime Eco Line Manual Vers 2014 06 34 analytikjena Problem Cause Corrective Actions Bands spreading outwards Diffusion of sample when loading Load samples as quick as possible and start the electrophoresis run a s a p after loading Diffusion of sample during run in the stacking gel Increase of stacking gel or increase current up to 25 for the run in the stacking gel Lower ionic strength of sample Match the ionic strength of the sample with that of the gel Bands are narrower than sample wells lonic strength of the sample is higher than ionic strength of the gel De salt the sample Use sample buffer of the same ionic strength as the gel Broad lanes at bottom of gel Samples with different buffers are loaded normal in gradient gels Use identical sample buffer for all samples loaded Skewed bands Gel not polymerised properly at the wells De gas gel solution before casting and or increase APS and TEMED concentration Control expiring date of the used solutions Salt concentration in the sample is too high De salt samples Separation gel is uneven at the top Overlay the gel carefully during polymerisation with bidist
4. Electrophoresis 33 Problem Cause Corrective Actions Leakage of gel solution Glass plates have not been aligned correctly Ensure that the two glass plates of the assembly are flush on a level surface Glass plates or spacers are dirty Clean thoroughly Uneven damaged edges of glass plates Use only glass plates that are not damaged Gasket of Casting Stand is dirty Clean thoroughly with a mild detergent and rinse with distilled water Dry without smearing Gasket of Casting Stand is damaged Replace gasket Electrophoresis Modul assembly is leaking Gaskets of the Electrophoresis Module are deformed dirty or too old Remove gaskets wash in warm water to remove excess of salts and place each gasket back into its groove If the gaskets are to old or brittle they have to be replaced Running time is too long Buffer is too much diluted Check buffer recipe and dilution Try again with fresh buffer Electrophoresis Modul assembly is leaking See above Running at too low current or voltage Make sure you are running using the suggested running conditions For running at const current the current value is per gel Running too fast Buffer is too concentrated Check buffer recipe and dilution Try again with fresh buffer If voltage is lower than usual when running at const current the buffer is probably too concentrated
5. 4 6 2 5 0 4 TEMED ul 21 21 21 21 21 10 APS ul 125 125 125 125 125 33 ml per 1 mm gel Percent Gel 7 5 10 0 12 5 15 0 17 5 30 0 8 Acrylamide Bis ml 80 108 137 165 19 3 1 88 M Tris HCl pH 8 8 ml 6 6 6 6 6 6 6 6 6 6 0 5 SDS ml 6 6 6 6 6 6 6 6 6 6 dist H2O ml 11 8 8 9 6 1 3 3 0 5 TEMED ul 28 28 28 28 28 10 APS ul 165 165 165 165 165 49 ml per 1 5 mm gel Percent Gel 75 10 0 12 5 15 0 17 5 30 0 8 Acrylamide Bis ml 11 9 161 203 245 28 7 1 88 M Tris HCl pH 8 8 ml 9 8 9 8 9 8 9 8 9 8 0 5 SDS ml 9 8 9 8 9 8 9 8 9 8 dist H2O ml 17 5 13 33 9 1 4 9 0 7 TEMED ul 42 42 42 42 42 10 APS ul 245 245 245 245 245 Eco Line Manual 2014 06 Biometra LT analytikjena x Stacking Gel 5 Eco Mini Gel Thickness 0 75 mm 1 0 mm 1 5mm Gel Volume 1 5 ml 2 ml 2 5 ml Acrylamide Bis 30 0 8 ml 0 2 0 3 0 4 0 625 M Tris HCl pH 6 8 ml 0 3 0 4 0 5 0 5 SDS ml 0 3 0 4 05 dist H2O ml 0 7 0 9 1 1 TEMED ul 2 2 3 10 APS ul 8 10 13 Eco Maxi Gel Thickness 0 75 mm 1 0 mm 1 5mm Gel Volume 2 5 ml 3 5 ml 5 0 ml Acrylamide Bis 30 0 8 ml 0 3 0 5 0 8 0 625 M Tris HCl pH 6 8 ml 0 5 0 7 1 0 0 5 SDS ml 0 5 0 7 1 0 dist H2O ml 1 2 1 6 2 2 TEMED ul 3 4 6 10 APS ul 13 18 26 A Attention The ammonium persulphate solution 10 should be prepared fresh every day and should be stored at 4 C refrigerator Eco Line Manual
6. 50686 11 e mail Service biometra com If you would like to send the unit back to us please read the following return instructions 11 1 Instructions for Return Shipment e Return only defective devices For technical problems which are not definitively recognisable as device faults please contact the Technical Service Department at Biometra Tel 49 0 5 51 50 88 1 10 or 12 Fax 49 0 5 51 50 88 1 11 e mail Service biometra com e Please contact our service department for providing a return authorization number RAN This number has to be applied clearly visible to the outer box Returns without the RAN will be not be accepted e Important Carefully clean all parts of the instrument from residues and of biologically dangerous chemical or radioactive contaminants If an instrument is contaminated Biometra will be forced to refuse to accept the device The sender of the repair order will be held liable for possible damages and losses resulting from insufficient decontamination of the device e Please prepare written confirmation use the Equipment Decontamination Declaration following on the next page that the device is free of biologically dangerous chemical or radioactive contaminants This confirmation must be attached to the outside of the packaging e Use the original packing or a similarly robust packing when returning the device If not available contact Biometra or your local distributor e Label the outside
7. Eco Line Overview Components included E 5 w 22 2 9 E 7 E g 5 8 88 2 apg 5 6 4 a Blea To g Q aw E ES g 5 373 8 G 2 2 o ga asa gt Aa 4 ika 6 wv no 4 nan a u x o CSB gt SS Os ZI ela ol 8 S se E 5 s S Ssg EFs ES ls 5 Item Order No 2 a AS DNS N Aw la DC Polyacrylamide Gel Electrophoresis Eco Mini System E 017 100 Eco Mini E 017 101 Eco Mini System EB 017 102 Eco Mini EB 017 103 Eco Mini System EBC with cooling option 017 104 Eco Mini EBC with cooling option 017 105 Eco Maxi System EB 017 400 Eco Maxi EB 017 401 Eco Maxi System EBC with cooling option 017 402 Eco Maxi EBC with cooling option 017 403 Tank Blot Apparatus Tankblot Eco Mini C with cooling option 018 100 Tankblot Eco Mini 018 101 Tankblot Eco Maxi C with cooling option 018 400 Tankblot Eco Maxi 018 401 Indication of buffer chamber E Electrophoresis EB Blotting EBC Cooling option Eco Line Manual Vers 2014 06 Ja analytikjena 10 Maintenance and Repair 10 1 Cleaning and Maintenance Rinse buffer tank bigfoot lid Electrophoresis Module Blot Module Blotting cassettes and Casting Stand thoroughly with running water and finally with distilled water after every use It is recommended to allow the chamber and Electrophoresis or Blotting Mod
8. Manual Vers 2014 06 12 analytikjena Eco Mini amp Tankblot Eco Mini Overview th a ra Eco Mini amp Tanblot Eco Mini System Overview Components included 4 v v E 4 3 2 2 a SE 3 le 7 w w ug p 2 8 E D 5 3 55353 2 po E a a 4 4 2 ER m g o vz S 4 E E ES 8 gt 2a p 5 2 bo f os sof 5 8 a5 2 G S 5 pe S 253 0 nos a a 3 5 V6 S E so z 5 z g2 gjg Mza 55 58 3 xE x 5 x lt 4 x gt gt Item Order No a a as a GD jni cn Ain VV jvj Eco Mini System E 017 100 Eco Mini E 017 101 Eco Mini System EB 017 102 Eco Mini EB 017 103 Eco Mini System EBC 017 104 Eco Mini EBC 017 105 Electrophoresis Module Eco Mini 017 175 Tankblot Eco Mini C 018 100 Tankblot Eco Mini 018 101 Blot Module Eco Mini 018 105 Eco Line Manual 2014 06 analytikjena ie Divider Plates for Eco Mini Electrophoresis Double gel capacity Divider Plate Set ind 2 combs Run up to 4 gels in a single run No additional Casting Stand and Electrophoresis Module required Fixed glass spacers for easy handling Spacer plate Divider Plate Notched plate Double gel sandwich for Eco Mini Eco Line Manual Vers 2014 06 14 Eco Maxi amp Tankblot Eco Maxi Overview analytikjena Eco Maxi amp Tanblot Eco Maxi System Overview Item Eco Maxi System EB Eco Maxi EB Ec
9. Overall size W x D x H Gel size W x L Blotting area W x L Gel compatibility Weight Chemical compatibility analytikjena Eco Mini approx 1 580 ml for electrophoresis buffer tank E approx 2 180 ml for electrophoresis buffer tank EB and EBC Approx 2 100 ml for blotting buffer tank EB and EBC Eco Maxi approx 5 400 ml for electrophoresis buffer tank EB and EBC approx 6 100 ml for blotting buffer tank EB and EBC Buffer tank EBC flow through Eco Mini 21 0 x 15 0 x 22 0 cm buffer tank E 21 0 x 15 0 x 25 0 cm buffer tank EB 21 0 x 15 0 x 26 5 cm buffer tank EBC Eco Maxi 32 5 x 15 0 x 38 5 cm buffer tank EB 32 5 x 15 0 x 40 0 cm buffer tank EBC Eco Mini 9 4 x 8 0 cm Eco Maxi 19 4 x 18 5 cm Eco Mini 9 4 x 8 0 cm Eco Maxi 22 0 x 19 0 cm Eco Mini hand cast and pre cast gels Eco Maxi hand cast gels Eco Mini max 2 5 kg depending on system Eco Maxi max 8 kg depending on system The components are not compatible with alcohol gt 10 e g ethanol or organic solvents e g acetone chloroform toluene benzene Use of organic solvents voids all warranties The instrument is designed for operation in closed laboratory rooms at ambient temperature from 5 C to 40 C and maximum relative humidity 80 for temperatures up to 31 C decreasing linearly to 50 relative humidity at 40 C Eco Line Manual 2014 06 Biometra INE analytikjena 7 1 3 Legal Notes Copyright All r
10. Vers 2014 06 7 analytikjena 7 2 Electrophoresis Buffer solutions SDS Sample Buffer for SDS PAGE gels under reducing conditions 2ml 0 625 M Tris HCl pH 6 8 0 2g SDS 5ml Glycerol 0 5 ml Mercaptoethanol 0 1 ml Bromophenol Blue 1 in ethanol 2 4 ml dist H2O A Attention Dilute the sample buffer 1 to 2 with the sample 1 vol buffer 1 vol sample Running Buffer pH 8 3 5 liter 15 1g Tris Base 72 0 g Glycine 5 0g SDS Running Buffer pH 8 3 10 liter 30 29 Tris Base 144 09 Glycine 10 0 g SDS Eco Line Manual 2014 06 analytikjena ya 7 3 Blotting Buffer Solutions Most transfer buffers for SDS PAGE gels are Tris Glycin buffers pH 8 3 At this pH the negatively charged proteins will move to the anode Using a transfer buffer with acidic pH will return the moving direction because the positively charged proteins will move to the cathode Typical transfer buffers are e 20 mM Tris 150 mM Glycine pH 8 3 e 7 5 mM Tris 1 2 mM Boric Acid pH 8 9 or e 25 mM Sodium Phosphate pH 6 5 Towbin buffer is the most commonly used buffer It works well for transfer over night 12 h at const 100 mA Towbin Buffer 20 25 mM Tris Base 150 192 mM Glycin 15 25 Methanol 0 02 0 1 SDS pH 8 1 8 5 The pH of Towbin buffer will not be adjusted and is dependent on the quality of the water and chemicals The buffer should be degassed prior to the addition of SDS and cooled to 4 C Higher conc
11. at the bottom e Tighten the Dummy plate in a similar way at the opposite side of the Electrophoresis Module in case of casting only a single gel The black Casting Stand gaskets must be clean and dry Do not soak the gaskets for prolonged periods prior to casting gels Take care that the black gaskets are exactly placed within the cavities in the stand and that they are inserted evenly Pull out both eccentric screws of the Casting Stand Eco Line Manual 2014 06 analytikjena 04 5 2 Place the Electrophoresis Module with the glass plates assemblies straight down into the Casting Stand Push the knob on each side of the Casting Stand with the flat bottom of the eccentric interlocking pin downwards Now simultaneously turn the eccentric screws on both sides of the Casting Stand for approx half a turn Note Don t turn the eccentric screw more than approx 180 Otherwise the glass plates can be shifted upwards and would get leaky Gel Casting Place a comb into the assembled gel cassette one comb per single gel sandwich 2 combs per double gel sandwich Mark the glass plate approx 5 to 10 mm below the comb teeth Remove the comb s and pour the running gel resolving gel up to this level Prepare the running gel monomer solution by adding all reagents except APS and TEMED Degas the solution under vacuum Do not use a sink water aspirator Add APS and TEMED to the degassed solution directly before pouring the
12. gel Use a glass or disposable plastic pipette and smoothly pour the solution up to the mark to prevent it from mixing with air Overlay the solution with bidistilled water or water saturated n butanol Prevent mixing Allow the gel to polymerise for approx 30 to 60 minutes Polymerisation is visible by the formation of a sharp dividing line between gel and overlay Remove the overlay solution from the surface of the polymerised gel If using water saturated n butanol rinse the gel surface with bidistilled water Do not leave the Eco Line Manual Vers 2014 06 22 analytikjena alcohol overlay on the gel for more than 1 h because this might dehydrate the top of the gel Prepare the stacking gel monomer solution by adding all reagents except APS and TEMED Degas the solution under vacuum Do not use a sink water aspirator Dry the top of the running gel with a sheet a filter paper before pouring the stacking gel Add APS and TEMED to the degassed solution directly before pouring the gel Use a glass or disposable plastic pipette and smoothly pour the solution until the top of the notched glass plate is reached Insert the desired comb between the two glass plates Allow the gel to polymerise for approx 30 to 60 minutes Gentle remove the comb after polymerisation and rinse the wells carefully with bidistilled water or running buffer System Assembly and Sample Loading After polymerisation lift the Electrophoresis Modul
13. lid for electrophoresis only Buffer tank EB for Eco Mini without lid for electrophoresis and blotting Buffer tank EBC for Eco Mini without lid for electrophoresis and blotting with cooling base Bigfoot Safety Lid for Eco Mini fits to all buffer tanks Electrophoresis Module for Eco Mini for 2 gels fits to all buffer tanks Glass plate for Eco Mini with fixed spacers 1mm Glass plate for Eco Mini with fixed spacers 0 75mm Glass plate for Eco Mini with fixed spacers 1 5mm Notched glass plate for Eco Mini Dummy plate for Eco Mini Divider Plate with fixed spacers 1 0 mm Divider Plate with fixed spacers 0 75 mm Divider Plate with fixed spacers 1 5 mm Divider Plate Set 2 divider Plates and 2 combs 10 well 1 0 mm Divider Plate Set 2 divider Plates and 2 combs 10 well 0 75 mm Divider Plate Set 2 divider Plates and 2 combs 10 well 1 5 mm Comb for Eco Mini 9 teeth 0 75mm MTP compatible Comb for Eco Mini 10 teeth 0 75mm Comb for Eco Mini 12 teeth 0 75mm Comb for Eco Mini 15 teeth 0 75mm Preparative comb for Eco Mini 1 well plus 2 marker wells 0 75mm Comb for Eco Mini 9 teeth 1 0mm MTP compatible Comb for Eco Mini 10 teeth 1 0mm MTP compatible Comb for Eco Mini 12 teeth 1 0mm MTP compatible Comb for Eco Mini 15 teeth 1 0mm MTP compatible Preparative comb for Eco Mini 1 well plus 2 marker wells 1 0mm Comb for Eco Mini 9 teeth 1 5mm MTP compatible Comb for Eco Mini 10 teeth 1 5mm
14. of the box with CAUTION SENSITIVE INSTRUMENT and the RAN number sticker Attach the Decontamination Declaration e Please enclose a note which contains the following a Sender s name and address b Name of a contact person for further inquiries with telephone number c Precise description of the fault which also reveals during which procedures the fault occurred if possible Eco Line Manual Vers 2014 06 46 analytikjena 12 Equipment Decontamination Certificate To enable us to comply with German law i e 71 StriSchV 17 GefStoffV and 19 ChemG and to avoid exposure to hazardous materials during handling or repair please complete this form prior to the equipment leaving your laboratory COMPANY INSTITUTE ADDRESS PHONE NO FAX NO E MAIL EQUIPMENT Model Serial No If on loan evaluation Start Date Finish Date Hazardous materials used with this equipment Method of cleaning decontamination The equipment has been cleaned and decontaminated NAME POSITION HEAD OF DIV DEP INSTITUTE COMPANY SIGNED DATE PLEASE RETURN THIS FORM TO BIOMETRA GMBH OR YOUR LOCAL BIOMETRA DISTRIBUTOR TOGETHER WITH THE EQUIPMENT PLEASE ATTACH THIS CERTIFICATE OUTSIDE THE PACKAGING INSTRUMENTS WITHOUT THIS CERTIFICATE ATTACHED WILL BE RETURNED TO SENDER General Information for Decontamination Eco Line Manual 2014 06 analytikjen
15. 0 68 6 0 Tel 49 0 5 51 50 68 6 10 or 12 Fax 49 0 5 51 50 68 6 66 Fax 49 05551 50 68 8 11 e mail Info biometra com e mail Service biometra com internet http www biometra de Eco Line Manual 2014 06
16. Biometra Eco Line Vertical Polyacrylamide Gel Electrophoresis amp Tank Blotting Apparatus Instruction Manual Model Eco Mini E System E 017 101 017 100 Eco Mini EB System EB 017 103 017 102 Eco Mini EBC System EBC 017 105 017 104 Eco Maxi EB System EB 017 400 017 401 Eco Maxi EBC System EBC 017 402 017 403 Tankblot Eco Mini C 018 100 Tankblot Eco Mini 018 101 Tankblot Eco Maxi C 018 400 Tankblot Eco Maxi 018 401 analytikjena Please read these instructions carefully before using this apparatus Biometra GmbH Service Department Rudolf Wissell Str 30 Rudolf Wissell Str 14 16 D 37079 Goettingen D 37079 Goettingen Tel 49 0 5 51 50 68 6 0 Tel 49 0 5 51 50 68 6 10 or 12 Fax 49 0 5 51 50 68 6 66 Fax 49 0 5 51 50 68 6 11 e mail Info biometra com e mail Service biometra com internet http www biometra de analytikjena This document describes the state at the time of publishing It needs not necessarily agree with future versions Subject to change Eco Line Manual 2014 06 analytikjena Contents 1 INIFOQUCHON crt nenne 3 11 Field of Applications esse een 3 12 Specifications ea 5 1 3 Legal Notes zesessiegerssengere eigenen 7 1 4 Meaning of the Instructions u a ee enieas ee itenetnne 7 2 Safety and Warning Notices uuuuunnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn 8 251 Definition of SM uses ee 8 2 2 General Safety Instruc
17. Eco Maxi Complete modular system with buffer chamber EB without cooling base Bigfoot Safety Lid 2 Blotting Cassettes colour coded each for 1 gel with size 22 cmx 19 cm 4 foam pads and manual Note buffer tank EB can be used with Electrophoresis Module 017 475 Eco Line Manual 2014 06 analytikjena re Accessories Eco Maxi 017 480 017 471 017 472 017 474 017 475 017 420 017 421 017 423 017 425 017 426 017 430 017 431 017 432 017 433 017 434 017 440 017 441 017 442 017 443 017 444 017 450 017 451 017 452 017 453 017 454 018 405 018 411 018 413 Casting Stand two place for Eco Maxi Buffer tank EB for Eco Maxi without bigfoot safety lid Buffer tank EBC for Eco Maxi without bigfoot safety lid Bigfoot Safety Lid for Eco Maxi fits to all buffer tanks Electrophoresis Module for Eco Maxi for 2 gels Glass plate for Eco Maxi with fixed spacers 1 mm Glass plate for Eco Maxi with fixed spacers 0 75 mm Glass plate for Eco Maxi with fixed spacers 1 5 mm Notched glass plate for Eco Maxi Dummy plate for Eco Maxi Comb for Eco Maxi 12 teeth 0 75 mm Comb for Eco Maxi 19 teeth 0 75 mm MTP compatible Comb for Eco Maxi 25 teeth 0 75 mm Comb for Eco Maxi 30 teeth 0 75 mm Preparative comb for Eco Maxi 1 well plus 2 marker wells 0 75 mm Comb for Eco Maxi 12 teeth 1 0 mm Comb for Eco Maxi 19 teeth 1 0 mm MTP compatible Comb for Eco Maxi 25 teeth 1 0 mm Comb
18. MTP compatible Comb for Eco Mini 12 teeth 1 5mm MTP compatible Comb for Eco Mini 15 teeth 1 5mm MTP compatible Preparative comb for Eco Mini 1 well plus 2 marker wells 1 5mm Blot Module for Tankblot Eco Mini Blotting cassette for Tankblot Eco Mini colour coded for 1 mini gel Foam pads for Tankblot Eco Mini 4 pcs for two cassettes Eco Line Manual Vers 2014 06 40 analytikjena Apparatus Eco Maxi 017 400 017 401 017 402 017 403 018 400 018 401 Eco Maxi System EB Complete modular system with buffer tank EB without cooling base Bigfoot Safety Lid Electrophoresis Module 2 glass plates with fixed 1 0 mm spacers 2 notched glass plates 2 combs 1 mm 12 wells Casting Stand and manual Note buffer tank EB can be used with Blot Module 018 405 Eco Maxi EB dto but without Casting Stand Eco Maxi System EBC Complete modular system with buffer chamber EBC with integrated cooling base Bigfoot Safety Lid Electrophoresis Module 2 glass plates with fixed 1 0 mm spacers 2 notched glass plates 2 combs 1 mm 12 wells Casting Stand and manual Eco Maxi EBC dto but without Casting Stand Tankblot Eco Maxi C Complete modular system with buffer chamber EBC with integrated cooling base Bigfoot Safety Lid 2 Blotting Cassettes colourcoded each for 1 gel with size 22 cm x 19 cm 4 foam pads and manual Note buffer tank EBC can be used with Electrophoresis Module 017 475 Tankblot
19. Maxi Transfer time for blotting with cooling should not be longer than 18 h at 30 W for Eco Mini and 18 h at 130 W for Eco Maxi using the cooling option Eco Line Manual 2014 06 Biometra INE max Temp analytikjena j 3 Setting Up Operation 3 1 Scope of Delivery Eco Line is available in multiple versions for electrophoresis and or blotting Eco Mini Complete system with buffer tank E EB or EBC Bigfoot Safety Lid with attached power cords Electrophoresis Module 2 sets of glass plates with fixed 1 mm spacers 1Dummy plate 2 combs 10 wells and manual Eco Mini System comes additionally with a Casting Stand for up to 2 single gel or double gel sandwiches Tankblot Eco Mini Complete modular system with buffer tank EB or EBC with cooling base Bigfoot Safety Lid with attached power cords Blot Module 4 Blotting Cassettes colour coded 8 foam pads and manual Eco Maxi Complete system with buffer tank EB or EBC Bigfoot Safety Lid with attached power cords Electrophoresis Module 2 sets of glass plates with fixed 1 mm spacers 2 combs 12 wells and manual Eco Maxi System comes additionally with a Casting Stand for up to 2 gels Tankblot Eco Maxi Complete modular system with buffer tank EB or EBC with cooling base Bigfoot Safety Lid with attached power cords Blot Module 2 Blotting Cassettes colour coded 4foam pads and manual Eco Line
20. Tanblot Eco Maxi System Overview Components included g 2 D 2 Ei g 2 Q K oO a Uc 3 amp E w w 3 g 7 E z a apo vy ie 3 29 F B 328 ajl z 8 5 58 8 22 23 3 153 YA 55 na S y a a 5 oO x 2 E z 5 5 2 KE S Ss 2 Ss 3 c 5 3 Sz te 3 xE x x 7 3 x lt gt Item Order No a as a D as AN N S a st Eco Maxi System EB 017 400 Eco Maxi EB 017 401 Eco Maxi System EBC 017 402 T 4 Eco Maxi EBC 017 403 Electrophoresis Module Eco Maxi 017 475 Tankblot Eco Maxi C 018 400 Tankblot Eco Maxi 018 401 Blot Module Eco Maxi 018 405 The Biometra Eco Line is designed for laboratory research only Eco Line Manual 2014 06 analytikjena 1 2 Specifications Construction Buffer Tank PMMA Bigfoot safety lid PMMA Blotting Module PMMA Blot cassette PMMA Electrophoresis Gel Module POM Casting Stand PE Combs Teflon Eco Mini combs Thickness Number of Wells Volume per Well mm I 2 x 25 and 385 2 x 35 and 515 43 45 35 25 2 x 55 and 780 65 70 55 37 Microtiter plate compatible multichannel pipet compatible Eco Maxi combs Thickness mm 2x 30 and 970 70 32 30 20 2x45 and 1 310 95 45 40 29 2x 75 and 2 340 165 75 70 52 Microtiter plate compatible multichannel pipet compatible Biometra ime Eco Line Manual Vers 2014 06 6 Buffer volume Cooling option
21. a ya Please contact your responsible health 8 safety officer for details Use of radioactive substances Please contact your responsible person for details Use of genetically change organism or parts of those Please contact your responsible person for details Eco Line Manual Vers 2014 06 Ja analytikjena 13 Note for Disposal of Electric Electronic Waste This symbol the crossed out wheelie bin means that this product should be brought to the return systems and or separate systems available to end users according to yours country regulations when this product has reached the end of its lifetime For details please contact your local distributor This symbol applies only to the countries within the EEA EEA European Economics Area comprising all EU members plus Norway Iceland and Liechtenstein Eco Line Manual 2014 06 analytikjena Ae 14 EC Declaration of Conformity EU Konformitatserklarung Goettingen January 2010 im Sinne der EG Richtlinie f r die elektromagnetische Vertr glichkeit 2004 108 EG following the EC directive about the electromagnetic compatibility 2004 108 EC Hiermit erkl ren wir dass folgende Elektrophorese und Blotting Systeme Herewith we declare that the following Electrophoresis and Blotting Systems Typ type Eco Mini E Eco Mini System E Eco Mini EB Eco Mini System EB Eco Mini EBC Eco Mini System EBC Eco Maxi EB Eco Maxi System EB Eco Maxi EBC Eco Max
22. d Never place the instrument on top of a Power Supply Make sure that the on off switch of the used external Power Supply is always free accessible Use only Biometra safety power cords for connecting the electrophoresis equipment to the Power Supply Eco Line Manual 2014 06 analytikjena 7 A A Danger High voltage The current to the cell provided from the external Power Supply enters the unit through the lid assembly providing a safety interlock to the user Current to the cell is broken when the lid is removed Do not attempt to circumvent this safety interlock and always turn Power Supply off before removing the lid or when working with the cell in any way Power to the instrument is supplied by an external DC voltage Power gt pply The output of this Power Supply must be isolated from external A _ und to issue that the DC voltage output floats with respect to ground A A A A might All Biometra Power Supplies meet this safety requirement Using the cooling option do not mix up the in and out plugs for the cooling water Best cooling is obtained using a refrigerated circulator chiller with a temperature of 5 C Attention Reduce flow rate to max 0 5 1 l min and use no organic solvents or alcohol Do not use alcohol e g methanol ethanol or organic solvents for cooling or cleaning the apparatus Fill the buffer tank with buffer up to the max fill line Us
23. d salts This prevents the gel to change size during the blotting transfer and reduces heating effects Cut blotting paper e g Whatman 3MM or GB005 to the size of the gel or the size of the cassette and soak 2 x 3 layers of thin 0 34 mm or 2 x 1 layer of thick approx 1mm with transfer buffer Wearing gloves cut the membrane to the size of the gel and blotting paper Mark the membrane to indicate the side that the samples will be on Wet the membrane according to the manufacturers recommendations followed by equilibration in transfer buffer for 15 minutes Soak 2 foam pads in transfer buffer Building up the gel sandwich is best done in a shallow plastic box Open the blotting cassette and place the cassette with the black side down on a flat surface Place a foam pad pre soaked with transfer buffer in the black half of the blotting cassette followed by 3 layers of thin or 1 layer of thick filter paper pre soaked with buffer Gently place the gel on top of the pre soaked filter paper Beginning at one end of the gel align the filter papers with the gel edge and slowly lower the other end driving out any bubbles Use a clean glass rod wetted in transfer buffer to roll out any trapped bubbles Carefully align and overlay the transfer membrane onto the gel in one smooth action Work from the centre and let the ends roll down Never re position the membrane as some transfer may occur on contact Use a clean glass rod wetted in tran
24. e Remove the blotting cassettes Eco Line Manual 2014 06 analytikjena 27 7 Buffers and Staining Destaining Solutions 7 1 Gel Solutions Running Gel Eco Mini 5 5 ml per 0 75 mm gel Percent Gel 75 10 0 12 5 15 0 17 5 30 0 8 Acrylamide Bis ml 1 4 1 8 2 3 2 8 3 2 1 88 M Tris HCI pH 8 8 ml 1 1 1 1 1 1 1 1 1 1 0 5 SDS ml 1 1 1 1 1 1 1 1 1 1 dist H2O ml 1 9 1 5 1 0 0 5 0 0 TEMED ul 5 5 5 5 5 10 APS ul 28 28 28 28 28 7 ml per 1 mm gel Percent Gel 75 10 0 12 5 15 0 17 5 30 0 8 Acrylamide Bis ml 1 7 2 3 2 9 3 5 4 1 1 88 M Tris HCl pH 8 8 ml 1 4 1 4 1 4 1 4 1 4 0 5 SDS ml 1 4 1 4 1 4 1 4 1 4 dist H2O ml 2 5 1 9 1 3 0 7 0 1 TEMED ul 6 6 6 6 6 10 APS ul 35 35 35 35 35 10 5 ml per 1 5 mm gel Percent Gel 7 5 10 0 12 5 15 0 17 5 30 0 8 Acrylamide Bis ml 2 6 3 5 44 5 3 6 1 1 88 M Tris HCl pH 8 8 ml 2 1 2 1 2 1 2 1 2 1 0 5 SDS ml 2 1 2 1 2 1 2 1 2 1 dist H2O ml 3 7 2 8 1 9 1 1 0 2 TEMED ul 9 9 9 9 9 10 APS ul 53 53 53 53 53 Eco Line Manual Vers 2014 06 analytikjena 28 Running Gel Eco Maxi 25 ml per 0 75 mm gel Percent Gel 75 10 0 12 5 15 0 17 5 30 0 8 Acrylamide Bis ml 6 1 8 2 10 4 125 14 6 1 88 M Tris HCl pH 8 8 ml 5 0 5 0 5 0 5 0 5 0 0 5 SDS ml 5 0 5 0 5 0 5 0 5 0 dist H2O ml 8 9 6 8
25. e for double gel sandwiches is approx 25 longer Eco Maxi const 15 25 mA per gel is recommended for the pre run and const 50 60 mA per Gel is recommended for the separation run The max Voltage has to be limited at 400 V Run time is depending on buffer and gel composition and should be approx 4 6 hours for SDS PAGE 5 5 System Reassembly and Gel Removal e After electrophoresis is completed turn off the Power Supply and disconnect the safety plugs e Remove the Bigfoot Safety Lid and slide out the Electrophoresis Module with the assemblies by carefully bending the Module towards the side with the Eco Line logo before lifting e Discard the running buffer e Loosen side clamps and remove gel assemblies Eco Line Manual Vers 2014 06 24 6 6 1 analytikjena Tankblotting Assembling the Gel Sandwich For gel sandwich assembly mounting of blotting cassettes inside the buffer tank EB or EBC and connecting to a power supply make sure that the membrane is placed at the side of the gel which is close to the anode positive electrode side red coloured if the target molecule are negatively charged Pre cool transfer buffer over night at 4 C After electrophoresis immediately remove the gel from the bottom plate Tip the plate up side down starting at one edge allow the gel to roll off into the transfer buffer Pre equilibrate the gel in cool transfer buffer for 15 minutes with gentle agitation to remove SDS an
26. e of lower buffer volume can generate higher buffer temperature and damage the instrument This products are designed and certified to meet EN 61010 1 safety standards Certified products are safe to use when operated in accordance with the instruction manual This instruments should not be modified or altered in any way Alteration of this instruments will void the warranty void the EN61010 1 certification and create a potential safety hazard Eco Line Manual Vers 2014 06 max V DC Electrophoresis Eco Mini Buffer tank E Buffer tank EB Buffer tank EBC Eco Maxi Buffer tank EB Buffer tank EBC Operating Conditions 200 200 200 400 400 max mA analytikjena max W 50 2 or 4 gels 10 50 C 50 2 or 4 gels 10 50 C 50 2 or 4 gels 10 50 C 125 2 gels 50 50 C 250 2 gels 100 50 C Max power rating for electrophoresis without cooling should not be longer A than 18h at 5 W gel for Eco Mini and 18 h at 25 W gel for Eco Maxi A Max power rating for electrophoresis with cooling should not be longer than 18 h at 5 W gel for Eco Mini and 18 h at 50 W gel for Eco Maxi Blotting Eco Mini Buffer tank EB Buffer tank EBC Eco Maxi Buffer tank EB Buffer tank EBC 100 100 60 100 300 300 1 000 1 000 30 50 C 30 50 C 60 50 C 130 50 C Transfer time for blotting without cooling should not be longer than 4 h at 30 W for Eco Mini and 6 h at 60 W for Eco
27. e with the gel assemblies from the Casting stand and slide into the selected buffer tank Fill the buffer tank with the running buffer to the indicated max level line Fill the chamber formed by the assemblies and the Electrophoresis Module with running buffer to just under the edge of the outer spacer glass plate approx 5 mm from the top of the gel assembly Make sure the buffer level is above the gap formed by the ears of the notched glass plate Dissolve the samples in sample buffer and carefully load into the wells of the stacking gel Set the bigfoot safety lid onto the unit Because of its asymmetric design this will only work in the proper orientation connecting the electrode of the buffer tank anode with the red power cord and the Electrophoresis Module cathode with the black power cord Eco Line Manual 2014 06 analytikjena za 5 4 Electrophoresis e Connect the safety plugs to a suitable Power Supply e g Standard Power Pack P25 or P25T e Be aware that the plus pole red is connected to the lower buffer reservoir e Apply power to the apparatus and begin electrophoresis Eco Mini const 10 mA per gel is recommended for the pre run and const 25 mA per Gel is recommended for the separation run The max Voltage has to be limited at 200 V Run time is depending on buffer and gel composition and should be approx 60 min to 120 min for SDS PAGE A Please notice Compared to single gel sandwiches the run tim
28. eeeeeeeeeeeseseenneeeeeeees 26 6 5 System Reassembly and Blot REmoOVal wwwwwwwmamuwimumimumamanunimim wawa 26 Eco Line Manual Vers 2014 06 5 analytikjena 7 Buffers and Staining Destaining Solutions uuunesssssnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn 27 71 ii ee ee see 27 1 2 Electrophoresis Buffer Solutions zu azi 30 7 3 Blotting Buffer Solutions nennen 31 7 4 Staining and Destaining Solutions uuuuussssssssssnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn nn 32 B Trouble Shooting nn 33 8 1 EIECHSPhNOLESIS ee 33 8 2 Tankbl llingsesseunnessensesesstnsekei2inkebeibuhbuehlilsbee 36 9 Product Information aes ates een EEE 38 10 Maintenance and Reparr unnnnne 44 10 1 Cleaning and Maintenance uuuuuusssssssnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn 44 10 2 gt gt AY Ya 1 gt IBEIEREREERTENREREEEEHNEERUEFERRUREREERERETTEEUETREREREEEEELTRTTERERUEREPEFREREETRUREEERTEEEERENEUE 44 10 3 Replacement of Spare Parts uuu 24444444444H0nnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn 44 TOA Other Ai ES iii ee 44 11 SEVIA nn 45 11 1 Instructions for Return Shipment usseneeeee ne 45 12 Equipment Decontamination CErtifiCAHte 222200 46 13 Note for Disposal of Electric Electronic Waste nnnnnnnnnnnnnnnnnnnnnnnnnnnnnn 48 14 EC Declaration Of Conformily uneussuninssien ann nina nenn ai 49 10 Wama Ve 50 Eco Line Manual 2014 06 analytikjena 3 1 Introduction 1 1 Fie
29. entration of SDS will increase transfer efficiency of proteins from the gel However this might lower binding capacity of the membrane A good buffer with higher ion strength for more quick transfer rates is 50 mM CAPS at pH 9 2 The pH of this buffer has to be adjusted with NaOH A very good buffer for quick transfer is 25 mM NasHPO at pH 9 0 0 1 This buffer should be used at const 300 mA A special transfer buffer for acidic gels with basic proteins IEF gels and urea gels is 0 7 Acetic Acid A Using acetic acid the transfer direction is reverse from to and transfer times are approx 50 shorter than with use of standard transfer buffers like Towbin Buffer A special transfer buffer for Glycoproteins Polysaccharides and Lipoproteins is 10 mM Sodium Borate pH 9 2 Eco Line Manual Vers 2014 06 as analytikjena 7 4 Staining and Destaining solutions Staining Solution 1 liter 2 09 Coomassie Brilliant Blue R 250 0 59 Coomassie Brilliant Blue G 250 425 ml Ethanol 50 ml Methanol 100 ml Acetic Acid 425 ml bidest Water A Attention Stir overnight filtrate before use store in dark bottle Destaining Solution fast destaining in 45 Ethanol 10 Acetic Acid 75 bidest Water slow destaining in 25 Isopropanol 10 Glacial Acid 45 bidest Water final destaining in 7 Acetic Acid in bidest water Eco Line Manual 2014 06 analytikjena 8 Trouble Shooting 8 1
30. for Eco Maxi 30 teeth 1 0 mm Preparative comb for Eco Maxi 1 well plus 2 marker wells 1 0 mm Comb for Eco Maxi 12 teeth 1 5 mm Comb for Eco Maxi 19 teeth 1 5 mm MTP compatible Comb for Eco Maxi 25 teeth 1 5 mm Comb for Eco Maxi 30 teeth 1 5 mm Preparative comb for Eco Maxi 1 well plus 2 marker wells 1 5 mm Blot Module for Tankblot Eco Maxi for up to 2 Blotting Cassettes each for 1 gel with size 22 cm x 19 cm and 4 foam pads Blotting cassette colour coded black red for Tankblot Eco Maxi Blot Module 1 pcs Foam pads for Tankblot Eco Maxi 4 pcs for two cassettes Eco Line Manual Vers 2014 06 42 analytikjena Blotting Paper 10 426 981 10 426 994 3017915 3030 931 Whatman GB005 200 mm x 200 mm 1 2 mm thick 25 sheets Whatman GB005 580 mm x 580 mm 1 2 mm thick 25 sheets Whatman 17Chr 460 mm x 570 mm 0 92 mm thick 100 sheets Whatman 3MM 580 mm x 680 mm cm 0 34 mm thick 100 sheets Nitrocellulose Blotting Membranes 10 401 396 10 401 196 10 401 180 10 402 580 10 439 196 Whatman Protran BA83 Pore size 0 20 um 30 cm x 3 m roll Whatman Protran BA85 Pore size 0 45 um 30 cm x 3 m roll Whatman Protran BA85 Pore size 0 45 um 30 cm x 60 cm 5 sheets pack Whatman Protran BA85 Pore size 0 45 um 33 cm x 56 cm 5 sheets pack Whatman Protran BA S 85 Pore size 0 45 um 30 cm x 3 m roll Eco Line Manual 2014 06 analytikjena 43 Overview
31. i System EBC Tankblot Eco Mini Tankblot Eco Mini C Tankblot Eco Maxi Tankblot Eco Maxi C Best Nr Order No 017 100 017 101 017 102 017 103 017 104 017 105 017 400 017 401 017 402 017 403 018 100 018 101 018 400 018 401 den grundlegenden Anforderungen der corresponds to the basic requirements of EG Niederspannungsrichtlinie 2006 95 EG entsprechen EC low voltage directive 2006 95 EC Juergen Otte PhD Quality Manager Eco Line Manual Vers 2014 06 ZA analytikjena 15 Warranty This laboratory instrument is produced with the highest practical standards of materials workmanship and design The design and manufacture of parts have been conceived with one purpose to produce units which will give satisfactory service Biometra GmbH guarantees this unit to be free from defects in materials or workmanship under normal use or service for 24 month from date of shipment If during this time this unit proves defective in materials or workmanship Biometra GmbH will repair or replace it free of charge if returned to us prepaid This guarantee does not cover damage in transit damage caused by carelessness misuse or neglect or unsatisfactory performance as a result of conditions beyond our control or consequential losses as a result of failure of our product Biometra GmbH Service Department Rudolf Wissell Str 30 Rudolf Wissell Str 14 16 D 37079 Goettingen D 37079 Goettingen Tel 49 0 5 51 5
32. ights reserved It is not allowed to copy and publish the manual or parts of it in any form as copies micro film or other methods without a written authorisation from Biometra Biometra is pointing out that applied company and brand names are usually protected trade marks Liability Biometra is not liable for damages and injuries caused by use not considering these operation instructions in parts or completely 1 4 Meaning of the Instructions Biometra recommends that you first read these instructions carefully Then assemble and disassemble the apparatus to become familiar with the system After these preliminary steps you should be ready to transfer a sample This operation instruction is part of the product and should be kept over the full life time of the instrument It should also be forwarded to subsequent owners and users Make sure that additions and updates are inserted into the operation instructions Eco Line Manual Vers 2014 06 2 2 1 N N gt PP gt gt analytikjena Safety and Warning Notices Definition of Symbols Symbol Definition Caution Refer to instruction manual Danger High voltage Fragile Using with direct current DC pb gt General Safety Instructions Delicate instruments Handle with care Do not operate this equipment if buffer or water leaks from the instrument if cracks are present in the body or the safety cover or ifthe electrical connection cables are worn or fraye
33. illed water or water saturated n butanol Make sure the Casting stand is levelled The Electrophoresis Module assembly is leaking through the gel and or along the side spacers Check that the gel is a solid gel inside the glass plates and check the setup of the apparatus to make sure that sealing with the gasket is perfect Check that spacer and com thickness are identical Streaked bands Sample overloading Use lower concentration for next run Sample precipitation Centrifuge sample before adding sample buffer and or use a lower acrylamide gel Frowning of outside lanes Leakage of buffer along the sides Ensure that the two glass plates of the assembly are flush on a level surface Eco Line Manual 2014 06 Biometra LT analytikjena 35 Problem Cause Corrective Actions Double bands Re oxidation or insufficient reduction of the sample If using a reducing reagent make sure that the buffer is not too old Increase the concentration of 2 mercapto ethanol or dithiothreitol DTT in the sample Biometra Ni Eco Line Manual Vers 2014 06 analytikjena 36 8 2 Tankblotting Problem Cause Corrective Actions No transfer Wrong order of membrane and gel Reverse order of membrane and gel gel on top of membrane Wrong connection to the Power Exchange connection Supply Red anode Black cathode
34. late 2 combs 10 wells Casting Stand and manual Note buffer tank EB can be used with Blot Module 018 105 Eco Mini EB dto but without Casting Stand Eco Mini System EBC Complete modular system with buffer tank EBC with cooling base Bigfoot Safety Lid Electrophoresis Module 2 sets of glass plates with fixed 1 mm spacers 1 Dummy plate 2 combs 10 wells Casting Stand and manual Note buffer tank EB can be used with Blot Module 018 105 Eco Mini EBC dto but without Casting Stand Tankblot Eco Mini C Complete modular system with buffer tank EBC with cooling base Bigfoot Safety Lid Blot Module 4 Blotting Cassettes colour coded 8 foam pads and manual Note buffer tank EBC can be used with Gel Module 017 175 Tankblot Eco Mini Complete modular system with buffer tank EB without cooling base bigfoot lid Blot Module 4 colour coded Blotting Cassettes 8 foam pads and manual Note buffer tank EBC can be used with Gel Module 017 175 Eco Line Manual 2014 06 analytikjena Accessories Eco Mini 017 180 017 169 017 170 017 171 017 172 017 175 017 120 017 121 017 122 017 125 017 127 017 183 017 184 017 185 017 187 017 188 017 189 017 130 017 131 017 132 017 133 017 134 017 140 017 141 017 142 017 143 017 144 017 150 017 151 017 152 017 153 017 154 018 105 018 111 018 113 Biometra Casting Stand two place for Eco Mini Buffer tank E for Eco Mini without
35. ld of Application The Biometra Eco Line is a modular tank style apparatus designed for the separation of proteins and nucleic acids DNA RNA in vertical polyacrylamide PAGE gels Interchangeable electrode modules allow electrophoresis and blotting using the same buffer tanks Using the Electrophoresis Module Gel Module the Eco Line runs up to 4 gels Eco Mini or up to two gels Eco Maxi The module accepts hand cast gels as well as pre cast gels Eco Mini only Using the Blot Module the apparatus allows blotting of one to two gels Eco Maxi or one to four gels Eco Mini simultaneously in individual cassettes Suitability Components included in the system Cooling Buffer tank Electrophoresis Blotting Eco Mini system E 017 100 Buffer tank see left bigfoot lid Eco Mini system EB A Electrophoresis Module 2 sets of glass plates 017 102 with fixed 1 mm spacers 1 dummy plate Eco Mini system EBC vIv v eBc 2 combs 10 wells casting stand and manual 017 104 en se G Buffer tank see left bigfoot lid Tankblot Eco Mini Blot Module 4 blotting cassettes colour coded 2 Pi 8 fiber pads and manual en lv emere 1 with addition of components required for blotting see furthermore for Tankblot Eco Mini systems with addition of components required for electrophoresis see furthermore for Eco Mini systems Eco Mini amp Tanblot Eco Mini System Overview Components included 2 T v E
36. n top of it e Double gel sandwiches Select a spacer plate and divider Plate of the desired gel thickness Place the Devider Plate on top of the spacer plate and the notched glass plate on top of the Divider Plate Spacer plate Divider Plate Notched plate e Prepare a second assembly if two gels will be needed e Slide the two glass plates assemblies single gel or double gel sandwiches or one assembly and one dummy plate into the Electrophoresis Module keeping the notched glass plate faced to the front of the module and the spacer plate faced to the wall of the buffer tank e Ensure that the glass plates of each assembly are flush on a level surface not in the Casting Stand A Leaking may occur if the plates are misaligned or damaged e Tighten side clamps of the Electrophoresis Module with the white screws before the Electrophoresis Module with the glass plate assemblies is placed into the Casting Stand In case that the clamps are too loose the gel assembly could get out of place during gel casting A Please take care that the screws are tightened alternating and strong enough Eco Line Manual Vers 2014 06 gt 0 analytikjena For Eco Mini there is only 1 screw per clamp For Eco Maxi there are 3 srews per clamp Make sure that the screws in the middle of each clamp are tightened first After this tighten the diametrically opposed screws crosswise e Check that all glass plates are flush
37. ng increase of voltage during transfer Blot sandwich dries out Reduce amperage soak blotting paper cuts with more transfer buffer Generation of air bubbles on the electrodes by electrolysis Chill buffers before use Reduce transfer time Insufficiently soaked transfer membrane Check procedure for prewetting membrane Eco Line Manual 2014 06 Biometra LT analytikjena 37 Problem Cause Corrective Actions Intensive heating up Amperage too high Reduce amperage Reduce transfer time Transfer time too long Chill buffers before use Reduce transfer time Conductivity of buffer too high Check transfer buffers Use buffers described in this manual Biometra Ni Eco Line Manual Vers 2014 06 38 9 analytikjena Product Information Apparatus Eco Mini 017 100 017 101 017 102 017 103 017 104 017 105 018 100 018 101 Eco Mini System E Complete system with buffer tank E Bigfoot Safety Lid Electrophoresis Module 2 sets of glass plates with fixed 1 mm spacers 1 Dummy plate 2 combs 10 wells Casting Stand and manual Note buffer tank E is not compatible with Blot Module 018 105 Eco Mini E dto but without Casting Stand Eco Mini System EB Complete modular system with buffer tank EB without cooling base Bigfoot Safety Lid Electrophoresis Module 2 sets of glass plates with fixed 1 mm spacers 1 Dummy p
38. o Maxi System EBC Eco Maxi EBC Electrophoresis Module Eco Maxi Order No 017 400 017 401 017 402 017 403 017 475 Buffer chamber EB Buffer chamber EBC with cooling option Bigfoot Safety Lid Electrophoresis Module Components included with fixed spacers 1 0 mm 2 x glass plates na a 2 T 4 a 2 y A 3 7 E D a To 2 v S 4 o 5 2 I D 2 a Ss e slz 5 S H 2 S G kai re a 2 gt gt 2 gt x lt gt N N 9 a Tankblot Eco Maxi C Tankblot Eco Maxi Blot Module Eco Maxi 018 400 018 401 018 405 Eco Line Manual 2014 06 analytikjena 3 2 Unpack and Check Unpack and carefully examine the Eco Line apparatus Examine for signs of damage and check the contents against the packing list Report any damage to BIOMETRA Do not attempt to operate this device if physical damage is present Please keep the original packing material for return shipment in case of service issues A I Attention A Please fill out and send back the warranty registration card This is important for you to claim full warranty 3 3 Installation Conditions Place the chamber in proximity to the Power Pack with which it is to be connected Be sure to place the chamber in a safe dry location away from the edge of the working surface 3 4 Connecting Conditions The Eco Mini Family apparatus have been designed to operate with D C current
39. p 2 o zS 3 g w amp Sy s Eg 5 3 a 533 we yg E 8 6 4 3 g p 28 Rn 2 3 s 4 E E EY B87 YY Soo S 2 6 Q 9 S c 2 a5 2 zZ gt 3 a a 23 2 no a a 3 5 o o oc ray A p 5 gt a 3 g 3e pa S 8 az g E 5 ge Ss 5 a S 52 5 5 we S 5 5 S 5 53 3 x 5 x 4 3 x gt Item Order No a a 858 0 sen Bi an Vi GB sim Eco Mini System E 017 100 Eco Mini E 017 101 Eco Mini System EB 017 102 Eco Mini EB 017 103 Eco Mini System EBC 017 104 Eco Mini EBC 017 105 Electrophoresis Module Eco Mini 017 175 Tankblot Eco Mini C 018 100 Tankblot Eco Mini 018 101 Blot Moduk Eco Mini 018 105 Eco Line Manual Vers 2014 06 P analytikjena Suitability Components included in the system Cooling Buffer tank Electrophoresis Blotting e IT Ten Buffer tank see left bigfoot lid oz 400 Electrophoresis Module 2 sets of glass plates Mes gt veel BAER with fixed 1 mm spacers 2 combs 12 wells 01 7 402 casting stand and manual Buffer tank see left bigfoot lid Tankblot Eco Maxi z Blot Module 2 blotting cassettes colour coded eaea 7171 ee Ferrer ana mena 1 with addition of components required for blotting see furthermore for Tankblot Eco Maxi systems with addition of components required for electrophoresis see furthermore for Eco Maxi systems Eco Maxi amp
40. sfer buffer to roll out any trapped bubbles Add 3 layers of thin or 1 layer of thick filter paper pre soaked with buffer on top of the membrane Complete the sandwich with a foam pad pre soaked in blotting buffer followed by the red half of the blotting cassette Eco Maxi Eco Mini Eco Line Manual 2014 06 6 2 6 3 analytikjena BE Insert the cassette into the Blotting Module Take care that the red side of the blotting cassette is located at the side of the Blotting Module with the red coloured electrode Eco Mini or at the side of the Blot Module with the red coloured screws for fixing the electrode Eco Maxi Up to 2 cassettes can be inserted into the Blotting Module of Eco Maxi and up to 4 cassettes can be inserted into the Blotting Module of Eco Mini Setting up the Buffer Tank EB or EBC Cool and degas the appropriate volume of transfer buffer approx 2 100 ml for use with Eco Mini or approx 6 100 ml for use with Eco Maxi Fill the buffer tank half to two third with transfer buffer Connect the cooling supply to the tank base connectors max 0 5 l min use hose clips to secure the tubing or place the system into a cold room Add a stir bar approx 5 cm in length to the tank to maintain uniform temperature and conductivity during the blotting transfer Adjust mixing device to max 200 rpm Slide the Blotting Module into the buffer tank Insert up to 4 cassettes into the Blotting Module of Eco Mini or insert
41. tions 222444440000n44nnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn 8 3 Setting Up Operation nennen 11 3 1 Scope of Delivery ZABEHRRERRREREREEEEEERDEREREHEREEREREREREEEEFEUEFREREREREREERRURFERETEREEREEREREFEFERFEREEN 11 32 Unpackand Check ensseees a EATE 15 33 Installation Conditions este een ua 15 34 Connecting Conditions u ea 15 4 COMPO iS iii casitienasencisnsctnsnnnstddenanisnniecesteisaanateseanesiatsannadanaaanntiviaanticxananasadensaaie 16 4 1 Systems Overview uuuessssnnsesnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn 16 4 2 Compatibility Overview 222400sunnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnennnnnnnnnnnn 18 5 Elebtroph resis aucnsn ann 19 5 1 Gel Cassette Assembly uuuussssnnsnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn 19 5 2 AGS gt 11 Ya ieii inia an a i iaaa haa 21 5 3 System Assembly and Sample Loading 444440sssnnss nn nennnnnnennnnnnn en 22 54 PIGCIODMORGSIG anne 23 5 5 System Reassembly and Gel Removal 244444000nnnnnsnnnnnnnnnnnnnnnnnnn nn 23 6 Tank UNG ende nn Eee ee 24 6 1 Assembling the Gel Sandwich uuus44424444444Bnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn 24 6 2 Setting up the Buffer Tank EB or EBC 22usssnnnnnnsssnnnnnnnnnnnnnnnnn nn 25 6 3 Transfer of more than 0n8 Gel 4 uu 2 22 2 25 6 4 Transfer Times and Conditions 0 ccccceeeeeeeeeeeeeeeeeeeecee
42. ule to air dry rather than drying with a towel to avoid damage of the electrode Glass plates and combs should be washed with a laboratory detergent and then rinsed thoroughly with distilled water Avoid extended submersion of spacer plates in water or cleaning solution Limit submersion to time used for cleaning Prolonged submersion compromises the integrity of the adhesive Do not use alcohol gt 10 e g methanol ethanol or organic solvents e g acetone for cleaning of electrodes or housing as this will cause acrylic to crack The system should never be autoclaved or placed in a microwave 10 2 Servicing Regular servicing is not necessary Nevertheless cleaning of the electrodes should be done using a damp cloth wetted with dest water after each transfer In case that the electrodes are clogged after longer time of use and the blotting transfer is inhomogeneous or reduced please contact the Biometra Service Department 10 3 Replacement of Spare Parts Only original spare parts mentioned in these operating instructions are allowed 10 4 Other Accessories See chapter 8 Product Informations for further details Eco Line Manual 2014 06 analytikjena HE 11 Service Should you have any problems with this unit please contact our service department or your local Biometra dealer Biometra GmbH Service Department Rudolf Wissell Stra e 14 16 D 37079 Goettingen Phone 49 0 5 51 50 686 10 or 12 Fax 49 05551
43. up to 2 cassettes into the Blotting Module and turn the locking handles on both sides downwards at Eco Maxi Add transfer buffer up to the marked level Place the Bigfoot Safety Lid on the buffer tank and connect the attached power cords to an appropriate power supply Start the blotting transfer Transfer of more than one Gel Transfer efficiencies might decrease if more than 1 blotting cassette is inserted In this case transfer times have to be increased to get the same transfer efficiencies as if blotting a single gel sandwich Eco Line Manual Vers 2014 06 sa analytikjena 6 4 Transfer Times and Conditions The best transfer time has to be determined experimentally There is no formula for determining transfer tine since there are too many variables involved to give specific transfer conditions To optimise transfer times it is useful to stain the gel as well as the membrane using a general staining method e g Coomassie blue or silver stain for protein gels and Ponceau red for proteins on membranes and to compare the results with an identical gel which has not been blotted 6 5 System Reassembly and Blot Removal e After electrophoresis is completed turn off the Power Supply and disconnect the safety plugs e Remove the Bigfoot Safety Lid and slide out the Blotting Module with the blotting cassettes by carefully bending the Module towards the side with the Eco Line logo before lifting e Discard the running buffer
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