manual - Isogen Life Science
Contents
1. Some possible solutions to potential problems are listed below If these suggestions are unclear or unsuccessful please contact PEQLAB Problem Agarose leaks into 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 PEGLAB Biotechnologie GmbH_v0507E 7 Instruction Manual PerfectBlue Horizontal Minigelsystems Problem Samples seem to 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 Problem Samples do not band sharply and appear diffuse in the gel Gels s2. 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 2 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 Minigelsystems 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 S
3. gels 7 x 8 cm W x 40 0708 Casting chamber Casting chamber for up to 3 gel trays 40 0708 CST Gel tray UV transmissible gel tray and gaskets 40 0708 UVT MultiCast Casting chamber Casting chamber and 3 UV transmissible gel trays 40 0708 MC Gaskets 2 rubber gaskets for gel tray 40 0708 GK Standard combs 1 5 mm 5 teeth 64 ur 40 0708 5D 1 5 mm 6 teeth 51 pl 40 0708 6D 1 5 mm 8 teeth 36 pl 40 0708 8D 1 5 mm 10 teeth 26 pl 40 0708 10D 1 5 mm 12 teeth 21 pl 40 0708 12D 1 0 mm 5 teeth 42 pl 40 0708 5C 1 0 mm 6 teeth 34 pl 40 0708 6C 1 0 mm 8 teeth 24 pl 40 0708 8C 1 0 mm 10 teeth 18 pl 40 0708 10C 1 0 mm 12 teeth 14 pl 40 0708 12C Preparative comb 1 5 mm 2 teeth 320 28 pl 40 0708 PD volumes are calculated for a gel thickness of 5 mm PerfectBlue Mini M Item Description Cat No Gel system Mini M complete system for gels 9 x 11 cm W x 40 0911 Casting chamber Casting chamber for up to 3 gel trays 40 091 1 CST Gel tray UV transmissible gel tray and gaskets 40 091 1 UVT MultiCast Casting chamber Casting chamber and 3 UV transmissible gel trays 40 091 1 MC Gaskets 2 rubber gaskets for gel tray 40 091 1 GK Standard combs 1 5 mm 5 teeth 86 pl 40 0911 5D 1 5 mm 8 teeth 51 pl 40 0911 8D 1 5mm 10 teeth 38 ur 40 0911 10D 1 5mm 12 teeth 30 pl 40 0911 12D 1 5 mm 14 teeth 25 pl 40 0911 14D 1 0 mm 5 teeth 58 pl 40 091 1 5C 1 0 mm 8 teeth 34 pl 40 091 1 8C 1 0 mm 10
4. model 2 combs 1 5 mm thick 12 and 20 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 Minigelsystems SYSTEM OVERVIEW The horizontal electrophoresis systems PerfectBlue Mini S M L and Mini L Revolution have been designed as all in one systems that make it possible to cast and run gels in the same chamber The user does not need any additional casting equipment such as grease agarose seals or other accessories to seal the gel tray for pouring the gel All PerfectBlue horizontal Minigelsystems include a UV transmissible gel tray which has the minimum of two comb positions allowing the user to run two sets of samples for equal distances simultaneously and a fluorescent ruler that helps in the precise photodocumentation of each gel run In total PEQLAB offers 6 different Minigelsystems In addition to the Mini S M L and L Revolution models that are described her
5. teeth 25 pl 40 0911 10C 1 0 mm 12 teeth 20 pl 40 0911 12C 1 0 mm 14 teeth 16 pl 40 0911 14C Microtiter combs 1 5 mm 9 teeth AO pl A0 0911 9D 1 5mm 18 teeth 16 pl 40 0911 18D 1 0 mm 9 teeth 27 pl 40 0911 9C 1 0 mm 18 teeth 11 pl 40 0911 18C Preparative comb 1 5 mm 2 teeth 439 28 pl 40 091 1 PD volumes are calculated for a gel thickness of 5 mm PEGLAB Biotechnologie GmbH_v0507E 9 Instruction Manual PerfectBlue Horizontal Minigelsystems PerfectBlue Mini L amp Mini L Revolution The same accessories are used for both models Mini Land Mini L Revolution Item Description Cat No Gel system Mini L complete system for gels 12 x 14 cm W x L 40 1214 Gelsystem Mini L Revolution complete system for gels 12 x 14 cm W x L 40 1214R Casing chamber Casting chamber for up to 3 gel trays 40 1214 CST Gel tray UV transmissible gel tray and gaskets 40 1214 UVT MultiCast Casting chamber Casting chamber and 3 UV transmissible gel trays 40 1214 MC Gaskets 2 rubber gaskets for gel tray 40 1214 GK Wall comb Wall comb for dividing up the gel tray 40 1214 WC Standard combs 1 5 mm 8 teeth 70 pl 40 1214 8D 1 5 mm 16 teeth 30 pl 40 1214 16D 1 5 mm 20 teeth 22 pl 40 1214 20D 1 5mm 24 teeth 17 pl 40 1214 24D 1 0 mm 8 teeth 47 pl 40 1214 8C 1 0 mm 16 teeth 20 pl 40 1214 16C 1 0 mm 20 teeth 15 pl 40 1214 20C 1 0 mm 24 teeth 11 pl 40 1214 24C Microtiter combs 1
6. 5 mm 9 teeth 40 ur A0 1214 9D 1 5 mm 12 teeth AO pl 40 1214 12D 1 5mm 25 teeth 16 pl 40 1214 25D 1 0 mm 9 teeth 27 pl 40 1214 9C 1 0 mm 12 teeth 27 ur 40 1214 12C 1 0 mm 25 teeth 11 pl 40 1214 25C Preparative comb 1 5 mm 2 teeth 596 28 pl 40 1214 PD volumes are calculated for a gel thickness of 5 mm JustCast adjustable casting chamber For the simple leak proof casting of up to three Mini S gels two Mini M gels two Mini L gels one Mini ExM gel or one Mini ExW gel Item Description Cat No JustCast Adjustable Casting Chamber for PerfectBlue 40 CST Minigelsystems including a 3 point levelling system with water level PEQLAB Biotechnologie GmbH_v0507E 10 Instruction Manual PerfectBlue Horizontal Minigelsystems 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
7. Instruction Manual PerfectBlue Horizontal Minigelsystems Mini S M L amp Mini L Revolution peer GmbH e 0507E Creating the future together Instruction Manual PerfectBlue Horizontal Minigelsystems 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 Mini S PerfectBlue Mini M PerfectBlue Mini L amp Mini L Revolution JustCast adjustable casting chamber Power Supplies Agaroses LITERATURE OO SON dd GO OG SE Go OO N N PEQLAB Biotechnologie GmbH_v0507E Instruction Manual PerfectBlue Horizontal Minigelsystems 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 retain all packaging materials until the delive
8. 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 HO 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 PEGLAB Biotechnologie GmbH_v0507E 5 Instruction Manual PerfectBlue Horizontal Minigelsystems 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 Model Gel size cm Gel thickness cm 0 25 0 5 0 75 1 0 PerfectBlue Mini S 7x8 Bxl 14 ml 28 ml 42 ml 56 ml PerfectBlue Mini M 9x11 BxL 25 ml 50 ml 75 ml 100 ml PerfectBlue Mini L 12x14 B xL 42 ml 84 ml 126 ml 168 ml PerfectBlue Mini L Revolution 12x 14 B x L 42 ml 84 ml 126 ml 168 ml The optimal rang
9. al 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 ml Glacial acetic acid 100 ml 0 5
10. e two wide format Minigelsystems are available Mini ExM and Mini ExW A comprehensive range of accessories is available for this range These include stand alone casting chambers for pouring up to 3 gels simultaneously while the chamber is in use the adjustable casting chamber JustCast a wide variety of standard combs microtiter combs preparative combs and wall combs that allow you to cast shorter gels in a standard gel tray For detailed information on available accessories visit www peglab de or see TECHNICAL SUPPORT AND ORDERING INFORMATIONS In contrast with all the other Minigelsystems the Mini L Revolution model is 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 Revolution Technology Technical properties PerfectBlue Cat No Gel size W xL Buffer volume Voltage Current Time required Mini S 40 0708 7x8cm 400 ml 20 150V 0 75 mA 30 60 min Mini M 40 0911 9x11cm 600 ml 20 150V 0 75 mA 45 90 min Mini L 40 1214 12x 14 cm 800 ml 20 150V 0 75 mA 60 120 min Mini L Revolution 40 1214R 12x14cm 1000 ml 20 150V 0 75 mA 60 240 min PEQLAB Biotechnologie GmbH_
11. e 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 migrate to the cathode during the electrophoresis leading to non homogeneous staining Improved results can be obtained by incubating the gel after the elec
12. ed 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 PEGLAB Biotechnologie GmbH_v0507E 8 Instruction Manual PerfectBlue Horizontal Minigelsystems 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 PEQLAB s products on www peglab deT PerfectBlue Mini S Item Description Cat No Gel system Mini complete system for
13. ge current see Technical properties 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 Minigelsystems 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 negatively charged in an alkaline to neutr
14. hould 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 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 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 ripp
15. ry 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 PerfectBlue Mini S Mini M Mini L and Mini L Revolution one buffer chamber with corrosion protected platinum electrodes one safety lid with attached power cords one UV transmissible gel tray with gaskets Mini S 2 combs 1 5 mm thick 6 and 10 teeth Mini M 2 combs 1 5 mm thick 10 and 14 teeth Mini L including Revolution
16. t the tray out of the chamber turn it 90 and replace it in the chamber with the first comb closest to the cathode side black electrode of the chamber The running position exposes the open ends of the agarose to the buffer 2 Pour enough compatible running buffer into the unit to fill chamber and completely cover and submerge the gel A Fill Line is located on each unit to clearly mark the correct buffer level See Technical properties for approximate buffer volumes needed for your unit Too little buffer may cause the gel to dry out during the run while excess buffer may slow DNA migration in the gel 3 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 4 Load prepared samples into the wells Samples should be mixed with a sample loading buffer giving weight to the samples so that they drop evenly into the wells and contain tracking dye to monitor the gel run 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 5 Carefully slide the lid with attached power cords onto the unit This will connect the power cords to the banana plugs to complete the circuit Plug the other end of the cords into an appropriate power supply 6 Turn on the power supply and run the gel at the appropriate volta
17. tandard 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 Minigelsystems NOTES PEQLAB Biotechnologie GmbH_v0507E Instruction Manual PerfectBlue Horizontal Minigelsystems 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 43 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 D peotob ger Creating the future together
18. trophoresis 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 PEGLAB Biotechnologie GmbH_v0507E 6 Instruction Manual PerfectBlue Horizontal Minigelsystems 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 estimated PEQLAB offers an extensive range of DNA and RNA markers For detailed information please contact us or visit www peqlab de TROUBLESHOOTING
19. v0507E 2 Instruction Manual PerfectBlue Horizontal Minigelsystems GENERAL INSTRUCTIONS Setting up the system and pouring the agarose gel i 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 shipping and convenient storage the gel tray is packaged inside the unit upon arrival NOTE This is also the correct 90 tray position for casting a gel To remove the gel tray hold the unit firmly with one hand grasp the long sides of the UVT gel tray and pull up slowly at an angle The tray needs to fit snug for leak proof gel casting so it may seem somewhat tight Walking the tray upwards at an angle may be helpful To cast a gel place the gel tray into the chamber so that the gasketed ends press against the walls of the buffer chamber Make sure the gel tray is pressed all the way down and rests level on the unit s platform 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 micro
20. wave oven until the agarose is completely dissolved The prepared gel must then 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 correct amount see Agarose Gel volumes and percentage of warm agarose lt 60 C onto the UVT gel tray that has been placed into the casting position in the gel box Immediately after pouring insert the desired comb or combs into the comb slots to form the sample wells Allow the gel to solidify completely A single comb may be placed in either groove on the gel tray If only a small portion of gel is required for proper sample separation then 2 combs may be used This also increases the number of samples per gel that may be run To conserve agarose a wall comb Model PerfectBlue Mini L only may be used to divide the gel tray in half Standard agarose should solidify completely in about 30 minutes If low melting point or a speciality agarose is used consult the instructions that came with the product PEGLAB Biotechnologie GmbH_v0507E 3 Instruction Manual PerfectBlue Horizontal Minigelsystems Loading of samples and electrophoresis 1 Once the gel is completely solidified lif
Download Pdf Manuals
Related Search