manual - Isogen Life Science
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1. Horizontal Maxigelystems Power Supplies Do not hesitate to contact us for advice on which Power Supply is most suitable for your application Item Ports max Voltage V max Current mA Power W Cat No EV222 3 200 200 20 55 EV222 EV243 3 400 300 50 55 EV243 EV231 4 300 1000 150 55 EV231 EV265 4 600 500 150 55 EV265 EV202 4 300 2000 300 55 EV202 EV261 4 600 1000 300 55 EV261 EV215 4 1200 500 300 55 EV215 EV232 4 3000 150 150 55 EV232 EV233 4 3000 300 300 55 EV233 EV262 4 6000 150 300 55 EV262 Agaroses Item Purpose Amount Cat No peqGOLD Universal Agarose Suitable for standard applications 100g 35 1010 Separation range between 0 05 and 50 kb 500g 35 1020 1000 g 35 1030 peqGOLD Universal Agarose Tabs Convenient tablet format Suitable for 50g 35 7010 standard applications Separation range 250g 35 7020 between 0 05 and 50 kb 500g 35 7030 peqGOLD Low Melt Agarose For the preparative separation of DNA 25g 35 2010 fragments between 0 08 and 20 kbp 100 g 35 2020 250 g 35 2030 peqGOLD MoSieve Agarose MS 500 Especially for high resolution separation of 25g 35 3010 small fragments 0 01 1 kbp 100g 35 3020 250 g 35 3030 peqGOLD MoSieve Agarose MS 1000 Especially for high resolution separation of 25g 35 4010 small fragments between 0 05 2 kbp 100g 35 4020 250 g 35 4030 pegGOLD MegaBase Agarose Especially for separation of larger DNA 25g 35 5010 fragments between 0 22. The larger size of the gel makes the need to cast and run a level gel very important for consistent reproducible results Level the unit using the thumbscrews on each side of the unit by slowly turning one thumbscrew at a time and lining up the bubble in the level with the center circle When preparing the gel use electrophoresis grade agarose and compatible electrophoresis buffer The gel may be prepared in various ways The percentage of agarose and the buffer used is determined by the size of the samples to be separated and further recovery of the samples see REQUIRED REAGENTS amp RECIPES The agarose and buffer are mixed and heated over a heat plate by stirring or in a microwave oven until the agarose is completely dissolved The prepared gel then must be cooled to below 60 C before casting to avoid warping the UVT gel tray due to excessive heat If numerous gels are to be run in one day a large volume of gel may be prepared and be placed in a covered bottle stored between 40 60 C in a water bath This provides a ready gel supply in a warm liquid form that will solidify quickly when gels are cast Pour or pipet the measured amount see Agarose Gel volumes and percentage of warm agarose lt 60 C onto the UVT gel tray that has been placed into the correct position in the gel box Immediately after pouring insert the desired comb or combs into the comb slots to form the sample wells If only a small portion of gel is required for
3. and 50 kbp 100g 35 5020 250g 35 5030 peqGOLD Pulsed Field Agarose Especially for Pulsed Field applications 25g 35 6010 100g 35 6020 250 g 35 6030 PEQLAB Biotechnologie GmbH_v0507E Instruction Manual PerfectBlue Horizontal Maxigelystems LITERATURE SAMBROOK J FRITSCH E F AND MANIATIS T 1989 Molecular Cloning A Laboratory Manual Cold Spring Harbor Laboratory Press NY FREDERIK M AUSUBEL et al Ed Short Protocols in Molecular Biology A Compendium of Methods from Current Protocols in Molecular Biology OGDEN R AND ADAMS D A 1987 Electrophoresis in Agarose and Acrylamide Gels Methods Enzymol 152 61 87 FOTADOR U SHAPIRO L E AND SURKS M I 1991 Simultaneous Use of Standard and Low Melting Agarose for the Separation and Isolation of DNA by Electrophoresis Bio Techniques 10 2 171 2 Boots S 1989 Gel Electrophoresis of DNA Anal Chem 61 8 551a 553a PEQLAB Biotechnologie GmbH_v0507E 12 Instruction Manual PerfectBlue Horizontal Maxigelystems NOTES PEQLAB Biotechnologie GmbH_v0507E Instruction Manual PerfectBlue Horizontal Maxigelystems Deutschland PEQLAB Biotechnologie GmbH Carl Thiersch Str 2b 91052 Erlangen Freecall D 0800 100 20 16 Tel 49 0 9131 610 70 20 Fax 49 0 9131 610 70 99 e mail info peglab de Internet www peglab de sterreich PEQLAB Biotechnologie GmbH Zweigniederlassung Linz Hafenstr 47 51 4020 Linz Tel 4
4. ml Glacial acetic acid 100 ml 0 5 M EDTA pH 8 0 made up to 1 using H O TBE Tris Borate EDTA Buffer 0 5 x working solution 45 mM Tris Borat 1 mM EDTA 5x stock solution 1 1 54 g Tris Base 27 5 g Boric acid 20 ml 0 5 M EDTA pH 8 0 made up to 1 using H O 0 5x TBE is sufficient for agarose gel electrophoresis For vertical electrophoresis in polyacrylamide gels 1x TBE is often applied due to the comparatively smaller buffer reservoirs of vertical electrophoresis chambers 5x TBE stock solutions tend to precipitate during long storage periods and should get remade Because of this property higher concentrations of TBE stock solutions should be avoided PEQLAB Biotechnologie GmbH_v0507E 5 Instruction Manual PerfectBlue Horizontal Maxigelystems Agarose Gel volumes and percentage PEQLAB offers an extensive range of high quality agaroses for many specific applications see TECHNICAL SUPPORT AND ORDERING INFORMATIONS The required volume of the gel is calculated using the following formula gel width cm x gel length cm x gel thickness cm required volume agarose solution ml The following volumes will result PerfectBlue Gel size cm Gel thickness cm 0 25 0 5 0 75 1 0 Maxi S 13 x 25 B xL 81 ml 162 ml 243 ml 325 ml Maxi M 20 x 25 B x L 125 ml 250 ml 375 ml 500 ml Maxi M Revolution 20 x 25 B x L 125 ml 250 ml 375 ml 500 ml Maxi L 23 x AO B x L 230 ml 460 ml 690 m
5. the chamber with the comb at the edge of the tray closest to the cathode side of the chamber Problem When the comb is removed from the gel the sample well is ripped and damaged Always make sure to allow the gel to solidify completely before moving the tray unit or removing the comb To avoid damage to the sample wells gently rock the comb back and forth lightly to loosen and then slowly pull the comb straight up out of the gel tray This rocking helps to avoid suction as the comb is removed Alternatively once casting is complete and the gel tray is placed in the running orientation simply submerge the gel in running buffer to help loosen the comb Problem The gel seems to run slower under the usual running conditions The volume of running buffer used to submerge the gel should only be between 3 5 mm over the gel surface Gel should be completely submerged to avoid the gel from drying out which can smear the bands and possibly melt the gel due to overheating If excessive running buffer is added the mobility of the DNA decreases and band distortion may result Excess buffer causes heat to build up and buffer condensation inside the unit may result PEQLAB Biotechnologie GmbH_v0507E 8 Instruction Manual PerfectBlue Horizontal Maxigelystems TECHNICAL SUPPORT AND ORDERING INFORMATIONS For technical questions please contact us by phone 49 0 9131 610 7020 or e mail info peglab de Please find detailed information on PEQLA
6. 0 teeth Maxi M Revolution 3 combs 1 5 mm thick 16 24 and 36 teeth Maxi L 4 combs 1 5 mm tick 2 each 25 and 50 teeth Maxi ExW including Revolution model 4 combs 1 5 mm thick 2 each 25 and 50 teeth User Manual SAFETY PRECAUTIONS Please read this Instruction Manual carefully before using the gel system Only use a CE marked DC power supply Always disconnect the gel system from the power supply before adding electrophoresis buffer Always disconnect the gel system from the power supply when it is not in use or before moving it Running conditions for this unit should not exceed the maximum operating voltage or current Do not fill the chamber with running buffer above the maximum fill line PEQLAB Biotechnologie GmbH_v0507E 1 Instruction Manual PerfectBlue Horizontal Maxigelystems SYSTEM OVERVIEW The horizontal electrophoresis systems PerfectBlue Maxi S Maxi M Revolution Maxi L and Maxi ExW Revolution are designed for the separation of medium to high sample numbers at long distances and provide flat even banding patterns and consistent results Their gel trays are getting sealed leak proof inside the chamber by using End Gates included in delivery The gasketed End Gates are specifically engineered to fit accurately into the gel tray grooves eliminating the need to tape or seal the tray before pouring gels Utilizing the built in levelling feature in all of the horizontal Maxig
7. 3 0 732 90 156 103 Fax 43 0 732 90 156 118 e mail info peglab at Internet www peglab at United Kingdom PEGLAB Lid 25 Barnes Wallis Road Fareham PO15 5TT Freecall UK 0808 20 21 302 Tel 44 0 1489 889 823 Fax 44 0 1489 660 040 e mail info peglab co uk Internet www pedlab co uk 4 peo ab ger Creating the future together
8. B s products on www peglab deT PerfectBlue Maxi S Item Description Cat No Gel system Maxi complete system for gels 13 x 25 cm W x L 41 1325 Gel tray UV transmissible gel tray and End Gates 41 1325 UVT Gaskets 2 rubber gaskets for End Gates 41 1325 GK End Gates 2 End Gates incl gaskets for gel tray sealing 41 1325 EG Wall comb Wall comb for dividing up the gel tray 41 1325 WC Standard combs 1 5 mm 8 teeth 78 pl 41 1325 8D 1 5 mm 12 teeth 49 pl A1 1325 12D 1 5 mm 16 teeth 34 pl A1 1325 16D 1 5 mm 20 teeth 25 pl A1 1325 20D 1 5 mm 24 teeth 20 pl A1 1325 24D 1 0 mm 8 teeth 52 pl 41 1325 8C 1 0 mm 12 teeth 33 pl 41 1325 12C 1 0 mm 16 teeth 23 pl A1 1325 16C 1 0 mm 20 teeth 17 pl 41 1325 20C 1 0 mm 24 teeth 13 pl 41 1325 24C Microtiter combs 1 5 mm 14 teeth 35 pl 41 1325 MTD 1 5 mm 28 teeth 16 pl 41 1325 28D 1 0 mm 14 teeth 25 pl A1 1325 MIC 1 0 mm 28 teeth 11 pl A1 1325 28C Preparative comb 1 5 mm 2 teeth 658 28 pl 41 1325 PD volumes are calculated for a gel thickness of 5 mm PerfectBlue Maxi M amp Maxi M Revolution The same accessories are used for the models Maxi M and Maxi M Revolution Item Description Cat No Gel system Maxi M complete system for gels 20 x 25 cm W x L 41 2025 Gelsystem Maxi M Revolution complete system for gels 20 x 25 cm W x L 41 2025R Gel tray UV transmissible
9. Instruction Manual PerfectBlue Horizontal Maxigelsystems Maxi S M L EBW Maxi M Revolution amp Maxi ExW Revolution peer GmbH e 0507E Creating the future together Instruction Manual PerfectBlue Horizontal Maxigelystems CONTENTS WARRANTY PACKAGING LIST SAFETY PRECAUTIONS SYSTEM OVERVIEW Technical properties GENERAL INSTRUCTIONS Setting up the system and pouring the agarose gel Loading of samples and electrophoresis Visualisation Cleaning REQUIRED REAGENTS amp RECIPES Electrophoresis buffers Agarose Gel volumes and percentage Ethidium bromide Loading buffer Sample buffer Molecular weight marker TROUBLESHOOTING TECHNICAL SUPPORT AND ORDERING INFORMATIONS PerfectBlue Maxi S PerfectBlue Maxi M amp Maxi M Revolution PerfectBlue Maxi L PerfectBlue Maxi ExW amp Maxi ExW Revolution Power Supplies Agaroses LITERATURE OO SON dd GO OG SE Go OO N N PEQLAB Biotechnologie GmbH_v0507E Instruction Manual PerfectBlue Horizontal Maxigelystems WARRANTY PEQLAB guarantees that the horizontal electrophoresis system you have received has been thoroughly tested and meets its published specification However immediately upon arrival please check carefully that the shipment is complete and has not been damaged in transit For missing parts or to report any kind of damage please contact PEQLAB see TECHNICAL SUPPORT AND ORDERING INFORMATIONS Please retai
10. e to the cathode during the electrophoresis leading to non homogeneous staining Improved results can be obtained by incubating the gel after the electrophoresis is finished in electrophoresis buffer containing 0 5 pg ml ethidium bromide for 5 to 20 min Subsequently the gel should get rinsed in electrophoresis buffer without ethidium bromide for up to 20 min in order to reduce background signal PEQLAB Biotechnologie GmbH_v0507E 6 Instruction Manual PerfectBlue Horizontal Maxigelystems Loading buffer Sample buffer Samples are prepared and mixed with loading buffer before applying to the prepared gel Sample buffers contain dyes for visibility and glycerol to provide weight to the samples This increased sample density ensures samples load evenly into the wells and do not float out during loading Dyes also migrate toward the anode end of the electrophoresis chamber at predictable rates allowing the gel run to be monitored In 0 5x TBE gels bromophenol blue migrates at the same rate as 300 bp DNA fragments and xylene cyanol approximately at the same rate as 4 kbp DNA fragments 6x DNA sample buffer 0 25 w v bromophenol blue 0 25 w v xylene cyanol FF 30 v v glycerol Molecular weight marker Markers are run on each gel to monitor the quality of sample separation and to enable a size estimation of specific bands By running a known marker of a specific concentration in parallel the DNA amount of the unknown samples can be est
11. elsystems allows the user to cast level gels quickly Four gel sizes are available to provide maximum versatility increasing sample capacity and varying sample running length Depending on the specific model the UV transmissible gel trays are equipped with 4 to 12 comb slots so that you can run multiple sample sets to equal distances simultaneously PEQLAB offers a wide variety of standard and microtiter combs in two different thicknesses 1 mm or 1 5 mm Microtiter combs allow for speedy sample loading using a multichannel pipette Furthermore preparative combs for large sample volumes and wall combs to cast gels in smaller sizes are available for increased versatility For deviding up the gel tray wall combs are put in the gel tray s slots instead of a common comb but should get sealed using agarose of higher percentage before casting the gel For detailed information on available accessories visit www peglab de or see TECHNICAL SUPPORT AND ORDERING INFORMATIONS The Maxi M Revolution and Maxi ExW Revolution models are equipped with an internal buffer recirculation system A trapping system captures hydrogen bubbles which are produced at the cathode due to electrolysis and directs them through an ascending tube to the opposing side of the buffer chamber where the anode is located During this hydrogen bubble migration the buffer circulates preventing the creation of detrimental pH or ion gradients Schematic drawing Revolutio
12. g Fill the chamber at the cathode end black electrode first 3 Then put the gel tray into the chamber remove the End Gates and completely cover and submerge the gel The running position of the tray exposes the open ends of the agarose to the buffer A Fill Line is located on each unit to clearly mark the correct buffer level Too little buffer may cause the gel to dry out during the run while excess buffer may slow DNA migration in the gel and band distortion 4 Carefully remove the comb or combs by tapping lightly to loosen and slowly lift straight up out of the gel tray to avoid damage to the wells 5 Load prepared samples into the wells Samples should be mixed with a sample loading buffer that gives weight to the samples so that they drop evenly into the wells and that contains tracking dyes to monitor the gel run Refer to TECHNICAL SUPPORT AND ORDERING INFORMATIONS for approximate well volumes NOTE It is wise to always run a sample lane of a known standard ladder to determine concentration and size of separated fragments after the gel run and to aid in photo documentation and analysis 6 Carefully slide the lid with attached power cords onto the unit This will connect the power cords to the banana plug to complete the circuit Plug other end of the power cords into an appropriate power supply 7 Turn on the power supply and run the gel at the appropriate current see Technical properties Carefully moni
13. gel tray and End Gates 41 2025 UVT Gaskets 2 rubber gaskets for End Gates 41 2025 GK End Gates 2 End Gates incl gaskets for gel tray sealing 41 2025 EG Wall comb Wall comb for dividing up the gel tray 41 2025 WC Standard combs 1 5 mm 8 teeth 127 pl 41 2025 8D 1 5 mm 12 teeth 82 pl 41 2025 12D 1 5 mm 16 teeth 59 pl A1 2025 16D 1 5 mm 20 teeth 45 pl A1 2025 20D 1 5 mm 24 teeth 37 pl 41 2025 24D 1 5 mm 28 teeth 28 pl 41 2025 28D 1 5 mm 32 teeth 22 pl 41 2025 32D 1 5 mm 36 teeth 20 pl A1 2025 36D PEQLAB Biotechnologie GmbH_v0507E 9 Instruction Manual PerfectBlue Horizontal Maxigelystems Standard combs continued 1 0 mm 8 teeth 85 pl 41 2025 8C 1 0 mm 12 teeth 54 pl 41 2025 12C 1 0 mm 16 teeth 39 pl 41 2025 16C 1 0 mm 20 teeth 30 pl 41 2025 20C 1 0 mm 24 teeth 24 pl 41 2025 24C 1 0 mm 28 teeth 19 pl 41 2025 28C 1 0 mm 32 teeth 15 pl 41 2025 32C 1 0 mm 36 teeth 13 pl 41 2025 36C Microtiter combs 1 5 mm 18 teeth AO pl 41 2025 18D 1 5 mm 21 teeth AO pl A1 2025 MID 1 5 mm 42 teeth 16 pl 41 2025 MT2D 1 0 mm 18 teeth 27 pl 41 2025 18C 1 0 mm 21 teeth 27 pl 41 2025 MIC 1 0 mm 42 teeth 11 pl 41 2025 MT2C Preparative combs 1 5 mm 2 teeth 1052 28 ul A1 2025 PD volumes are calculated for a gel thickness of 5 mm PerfectBlue Maxi L Item Description Cat No Gel system Maxi L complete system for gels 23 x 40 cm W x L 41 2340 Gel t
14. ie GmbH_v0507E 7 Instruction Manual PerfectBlue Horizontal Maxigelystems Problem Samples do not band sharply and appear diffuse in the gel Gels should be no more than 5 mm thick and be allowed to solidify completely before running Standard agarose should solidify in about 30 minutes If low melting point agarose is used it may be necessary to completely solidify gels at a cooler temperature in the refrigerator or cold room Gels should be submerged in 3 5 mm of buffer to avoid gel dry out but excess buffer gt 5 mm can cause decreased DNA mobility and band distortion Problem Samples are not moving as expected through the gel remaining in the wells running backwards or diffusing into the gel Check to be sure that a complete power circuit is achieved between the unit and the power supply Platinum wire and banana plugs should be intact To test simply fill the unit with running buffer and attach to the power supply without a gel or gel tray in the unit The platinum wires on both sides of the unit should produce small bubbles as the current passes through If a complete circuit does not exist there will be little to no bubbles If samples appear to run backwards through the gel or there are no bands visible check to be sure that the gel tray was placed in the electrophoresis chamber in the proper orientation If the orientation or polarity is reversed the samples will run backwards or migrate off the gel The tray should be placed in
15. imated PEQLAB offers an extensive range of DNA and RNA markers For detailed information please contact us or visit www peqlab de TROUBLESHOOTING Some possible solutions to potential problems are listed below If these suggestions are unclear or unsuccessful please contact PEQLAB Problem Agarose leaks into casting chamber when pouring gel Check to see if the gasket is firmly seated in the grooves on the ends of the UVT gel tray Reseat gasket if necessary by removing and rinsing under warm running water then reseat evenly in the tray groove Problem Bands seem to be running at an angle Gel smiling Check to be sure the casting is being done on a level surface Also confirm that the gel tray is inserted all the way into the unit and rests on the platform for level gel casting The voltage may be too high Try lowering the voltage setting on the power supply Problem Samples seem fo be running unevenly in certain areas Check that the platinum electrode wire is intact and running evenly across the base of the chamber and up the side to the junction of the banana plug If there appears to be a break in the electrode connection contact PEQLAB immediately This problem may also be caused by regularly casting with very hot agarose gel gt 60 C Always cool the melted agarose to below 60 C before casting to avoid warping the UVT gel tray Warping the gel tray will cause all subsequent gels to be cast unevenly PEQLAB Biotechnolog
16. l 920 ml Maxi ExW 23 x 25 B x L 144 ml 288 ml 432 ml 575 ml Maxi ExW Revolution 23 x 25 B x L 144 ml 288 ml 432 ml 575 ml The optimal range of DNA fragment sizes separated by any electrophoresis experiment is dependent on the agarose concentration of the gel The higher the agarose concentration the better small fragments are separated from each other and vice versa However for the smallest or largest fragment lengths the usage of specialized agaroses or polyacrylamide gels should be considered see table below since a 3 agarose solution solidifies rapidly and a 0 3 agarose gel is very soft and difficult to handle Agarose content w v Agarose g Puffer ml optimal separation range kb 0 3 0 3 100 5 30 0 5 0 5 100 1 15 0 7 0 7 100 0 8 10 1 0 1 0 100 0 5 7 1 2 1 2 100 0 3 6 1 5 1 5 100 0 2 4 2 0 2 0 100 0 1 3 3 0 3 0 100 lt 0 1 Ethidium bromide The gel may be stained during or following the run with a variety of stains for photodocumentation The most common stain for DNA is ethidium bromide Because of its capacity to intercalate between the bases of a nucleic acid strand and altering the sterical properties of DNA ethidium bromide is judged to be highly mutagenic Therefore appropriate safety measures must be applied Ethidium bromide may be added directly to the gel before pouring it at a concentration of 0 1 to 0 5 pg ml However being positively charged ethidium bromide will migrat
17. n Technology Technical properties PerfectBlue Cat No Gel size W xL Buffer volume Voltage Current Time required Maxi S 41 1325 13 x 25 cm 1600 ml 20 250 V 0 150 mA 60 120 min Maxi M 41 2025 20 x 25 cm 2300 ml 20 250 V 0 150 mA 60 240 min Maxi M Revolution 41 2025R 20 x 25 cm 2000 ml 20 250V 0 150mA 60 360 min Maxi L 41 2340 23 x 40 cm 4500 ml 20 250 V 0 150 mA 60 360 min Maxi ExW 41 2325 23 x 25 cm 3000 ml 20 250 V 0 150 mA 60 240 min Maxi ExW Revolution 41 2325R 23 x 25 cm 2800 ml 20 250 V 0 150 mA 60 360 min PEQLAB Biotechnologie GmbH_v0507E Instruction Manual PerfectBlue Horizontal Maxigelystems GENERAL INSTRUCTIONS Setting up the system and pouring the agarose gel l Remove the lid from the gel box by holding the front of the buffer chamber with one hand and pulling the lid off by holding the center of the back of the lid The cover is attached to the back of the unit at the connection of the power cords to the banana plugs For convenient storage the gel tray should be placed inside the unit on the platform with the gasketed End Gates in position Lift the casting tray out of the buffer chamber To cast a gel place the gel tray into the chamber making sure the gel tray rests level and is centered on the platform Slide the gasketed End Gates into the outermost grooves on either side of the gel tray The End Gates should be inserted tightly into the grooves with the gasket side facing out
18. n all packaging materials until the delivery has been completely checked since this will speed up the return of goods if required and reduce environmental impact Any form of returns replacements or credit notes must be agreed in advance by PEQLAB For the complete range of PerfectBlue electrophoresis and blotting systems PEQLAB guarantees a warranty period of 36 months if the products have been used solely according to the instruction manual and if not agreed differently After the warranty period has expired PEQLAB can offer repairs at low costs No liability is accepted for loss or damage arising from incorrect use PEQLAB s liability is limited to the repair or replacement of the unit or refund of the purchase price at PEQLAB s discretion PEQLAB is not liable for any consequential damages PEQLAB reserves the right to alter the technical specifications of the PerfectBlue electrophoresis or blotting systems without prior notice This will enable us to implement developments as soon as they arise PACKAGING LIST Unless differently agreed or marked on the delivery note the following items are included in shipment for the models Maxi S Maxi M Maxi M Revolution Maxi L Maxi ExW and Maxi ExW Revolution one buffer chamber with corrosion protected platinum electrodes one safety lid with attached power cords one UV transmissible gel tray and two End Gates one casting chamber Maxi S 3 combs 1 5 mm thick 12 16 and 2
19. proper sample separation multiple combs may be used to run 2 3 4 5 or 10 sets depending on the model of samples for equal distances simultaneously increasing the number of samples that may be run per gel To conserve agarose a wall comb may also be used to divide and use just half the length of the gel tray If a wall comb is used pipet a bead of agarose along the bottom and side edges of the wall comb once it has been placed in the tray to seal the combs edges to the trays bottom and sides Once this bead is solidified the cooled gel may be poured as described Alternately regular tape cut slightly longer then the comb can be placed flat along the comb s surface and the comb angled into place in the gel tray Extra tape is then placed on the outside of the comb in the excess tray area to reinforce the corners NOTE Allow the gel to solidify completely Standard agarose should solidify completely in about 30 minutes If low melting agarose or a speciality agarose is used please consult the instructions that came with the product PEQLAB Biotechnologie GmbH_v0507E 3 Instruction Manual PerfectBlue Horizontal Maxigelystems Loading of samples and electrophoresis 1 Once the gel is completely solidified carefully lift the tray out of the chamber as described above and place it beside the unit 2 Pour enough compatible running buffer into the unit to fill both compartments and allow it to stand for about 15 minutes prior to runnin
20. ray UV transmissible gel tray and End Gates 41 2340 UVT Gaskets 2 rubber gaskets for End Gates 41 2340 GK End Gates 2 End Gates incl gaskets for gel tray sealing 41 2340 EG Wall comb Wall comb for dividing up the gel tray 40 2314 WC Microtiter combs 1 5 mm 25 teeth AO pl 40 2314 MTD 1 5 mm 26 teeth AO pl 40 2314 26D 1 5 mm 50 teeth 16 pl 40 2314 MT2D 1 0 mm 25 teeth 27 pl 40 2314 MTC 1 0 mm 26 teeth 27 pl 40 2314 26C 1 0 mm 50 teeth 11 pl 40 2314 MT2C volumes are calculated for a gel thickness of 5 mm PerfectBlue Maxi ExW amp Maxi ExW Revolution The same accessories are used for the models Maxi ExW and Maxi ExW Revolution Item Description Cat No Gelsystem Maxi ExW complete system for gels 23 x 25 cm W x L 41 2325 Gelsystem Maxi ExW Revolution complete system for gels 23 x 25 cm W x L 41 2325R Gel tray UV transmissible gel tray and End Gates 41 2325 UVT Gaskets 2 rubber gaskets for End Gates 41 2340 GK End Gates 2 End Gates incl gaskets for gel tray sealing 41 2340 EG Wall comb Wall comb for dividing up the gel tray 40 2314 WC Microtiter combs 1 5 mm 25 teeth AO pl 40 231 4 MTD 1 5 mm 26 teeth AO pl 40 231 4 26D 1 5 mm 50 teeth 16 pl 40 231 4 MT2D 1 0 mm 25 teeth 27 ul 40 2314 MTC 1 0 mm 26 teeth 27 pl 40 2314 26C 1 0 mm 50 teeth 11 pl 40 231 4 MT2C volumes are calculated for a gel thickness of 5 mm PEQLAB Biotechnologie GmbH_v0507E 10 Instruction Manual PerfectBlue
21. tor the gel run to avoid samples running into the path of another set of samples Visualisation When the gel run is completed and the tracking dye has migrated as far through the gel as desired or to the end of the gel turn off the power supply and slide off the lid to disconnect from the power source Carefully remove the tray containing the gel wear gloves if ethidium bromide is present The UV transmissible gel tray makes for simple visualisation and photography with a UV light source without the need to remove the gel from the tray The gel tray may be placed back into the casting chamber for convenient transport to the darkroom and to avoid damage to the gel Cleaning The buffer chamber and tray should be rinsed under warm running water after each use Use a mild detergent to get rid of any debris It is recommended to allow the chamber to air dry rather than drying with a towel to avoid damage to the electrode wires Do not use ethanol or other organic solvents to clean acrylic products because organic solvents cause acrylic to craze or crack PEQLAB Biotechnologie GmbH_v0507E 4 Instruction Manual PerfectBlue Horizontal Maxigelystems REQUIRED REAGENTS amp RECIPES Electrophoresis buffers In general electrophoresis buffers supply the ions necessary for electrophoresis and establishing a certain pH value in which the target molecule adapts to its the required electric charge Nucleic acids for example will be negati
22. vely charged in an alkaline to neutral surrounding Additionally electrophoresis buffers often contain reagents which protect the target molecule from degradation e g EDTA which complexes bivalent cations and therefore inhibits DNases If electrophoresis under denaturing conditions is desired like for the electrophoresis of RNA electrophoresis buffers will additionally contain reagents that eliminate the formation of secondary structures Below you will find recipes for TAE and TBE two of the most commonly used buffers for the electrophoresis of DNA If the intention is to eventually isolate DNA from the gel TAE buffer should be chosen In comparison to TBE migration will be faster and a better resolution of supercoiled DNA will be achieved when using TAE However because of TAE s limited buffering capacity TBE should be selected for performing extended electrophoresis separations and if the electrophoresis chamber does not possess a system for buffer recirculation PEQLAB s PerfectBlue Revolution Systems are equipped with such an internal buffer recirculation system which effectively prevents the formation of pH and ion gradients during extended runs Since agarose tends to create finer pore sizes and a more solid matrix in TBE diffusion of DNA will be reduced and a more discrete band pattern will be achieved TAE Tris Acetate EDTA Buffer 1x working solution AO mM Tris acetate 1 mM EDTA 50x stock solution 1 242 g Tris Base 57 1
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