S4S and S3S Aluminum Backed Sequencer User Manual
Contents
1. spillage with 0 2mm with 0 4mm Spacers Spacers S38 35 x 45cm 70m SAS 20 x 45cm 50ml These volumes give you extra in case of spillage 2 2 Aluminum Sequencer Thermo Scientific Section 2 Setting Up Casting co ntinued 6 Fill the cassette resting at a slight incline If a regular test tube rack is being used place the plate assembly on the lab bench with the upper plate edges resting on the test tube rack This provides about 1 to 3 inches of elevation The solution may be poured directly from a beaker or flask or introduced using a pipette or syringe Pour gel to the top of the shorter glass plate 7 Sharktooth comb Carefully insert the flat edge of the sharktooth comb between the plates to a depth of 2 to 3mm below the shorter plate If the flat edge of the comb is inserted too deeply more than 4 or 5mm during gel casting the resulting trough will be too deep to load samples easily with a pipette It is wise to make marks on the comb to distinguish the right left sides so the comb can be put back in the same orientation for sample loading later in the event that the material has any slight variation in thickness Place binder clamps over the comb and glass plates along the upper edge to force the glass plates against the comb This will ensure a tight fit of the comb during and after polymerization If top binder clamps are not used the comb may not be tight enough to prevent sample leaks between wells following2. SCIENTIFIC
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4. loading samples Combs and spacers ideally should be stored in sets with each other and labeled with their thickness 1 Lay the larger blank glass plate suspended on a flat support on the laboratory bench A regular test tube rack works nicely 2 Place the two side spacers on the long edges of the glass being careful not to touch the cleaned plate The third bottom spacer can then be placed on the bottom of the glass 3 Lay the shorter offset or notched glass plate on top and line up the glass bottom and sides 4 If using casting tape tape all three sides of the cassettes for a leak proof seal Care must be taken to eliminate any air channels by pulling the tape firmly and pressing the tape onto the plates making hospital EV corners at the edge Binder clamps e may be used instead of casting gt E w tape or used in addition to tape to gt ensure a leak proof seal When I iq binder clamps are used a third i R bottom spacer must also be KH included Binder clamps are placed EE over thespacon being Figure 2 2 Clamps instead of Tape careful not to overlap the gel as this may alter the gel thickness causing thin spots on the gel edges 5 Prepare the appropriate acrylamide gel for your experimental technique in a suitable volume Regular standard size sequencing plates of 35 x 45cm with 0 4mm spacers require about 70 milliliters of solution Prepare a volume that is slightly larger to compensate for leaking or
5. oth de ewe cos seme ee oes 2 1 Casting with Casting Tape Binding Clamps 2 2 Loading Samples sic stern Doct teats da ee 2 4 Pre Runnine the Gel tai EE 2 4 Using the System e su ENEE ra a an 3 1 Running the Gel ascesa aope Rees ROS eee Ne AD 3 1 Technical TIPS da co tied AG ln al 4 1 Loading and Sequencing escocesa 4 Difference between Well and Sharktooth Comb 4 2 The Third Spacer vaccine ye tee e gee Gee ches 4 2 The Difference between Notched and Offset Glass 4 2 Troubleshooting iia 5 1 Care and Cleaning 00ccccee cece cece eee eeaeees 6 1 CAPA a ae eee ee 6 1 Tips About Caring for Your System cuicos 1 2a eins 6 3 Aluminum Sequencer V Section 1 General Information The Aluminum Backed Sequencing System provides smile free high resolution sequencing gels The floating aluminum plate allows contact with the entire surface of the glass plates providing a leak free seal and uniform heat distribution without overtightening the clamps The system features a safety interlocking lid and removable lower buffer chamber for improved safety The upper buffer chamber drains into a removable container on the back for easy buffer disposal and storage Unpack and Check Before starting unpack the unit and inventory your order If any parts are Your Order Missing contact Technical Services within 7 days of purchase Reference the order or catalog number on your invoice and ch
6. pre heated buffer to the upper buffer chamber a few times during the run to prevent this Aluminum Sequencer 5 1 Section 5 Troubleshooting 5 2 Aluminum Sequencer PROBLEM Buffer is leaking from between the UBC and the glass plates SOLUTION Check the red silicone gasket to make sure it is firmly seated Grease the gaskets if necessary If a gasket is old it may need to be replaced See the catalog for part numbers PROBLEM Sparks and smoke are coming from the unit SOLUTION First turn off the power to the apparatus This is usually caused by a slow leak from the upper chamber Do not run the apparatus under these conditions Call Technical Services if this occurs PROBLEM Bands are smearing SOLUTION Smearing of bands may be caused by template DNA that is not pure Try repurifying the template An old gel or the use of old gel components may also give poor quality results Try making up new gel solutions DNA samples that were not heat denatured prior to gel loading will not run as flat smooth bands Always remember to heat denature samples 5 minutes at 95 C before loading PROBLEM Blurring of bands SOLUTION Poor contact between the gel and X ray film during autoradiography will cause the bands to appear blurred and reading the data will be difficult This can be eliminated by adding weight to the film cassette during exposure When using S3S if the area is not completely rinsed from the gel before X ray fi
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8. Aluminum Backed Sequencer Models S4S and S3S Operating and Maintenance Manual 7007348 Rev 0 Visit us online to register your warranty www thermoscientific com warranty amp Owl Preface MANUAL NUMBER 7007348 0 4 26 12 Transfer to Marietta was T rex 01 2004 ccs REV ECR ECN DATE DESCRIPTION By Thermo Scientific Aluminum Sequencer i Preface CAUTION Contains Parts and Assemblies Susceptible to Damage by Electrostatic Discharge ESD Important Read this instruction manual Failure to read understand and follow the instructions in this manual may result in damage to the unit injury to operating personnel and poor equipment performance A Caution All internal adjustments and maintenance must be performed by qualified service personnel A Warning To avoid the risk of personal shock always disconnect the gel box from the power supply Further the power supply must be equipped with a shut down ondisconnect circuit Do not move the unit unless the power source to the unit has been disconnected 4 Statement of Proper Use Use this product only for its intended purpose as described in this manual Do not use this product if the power leads are damaged or if any of its surfaces are cracked Running conditions for this unit should not exceed the name plate readings found on the side panel This system is designed to meet IEC 1010 1 safety standards IEC 1010 1 is an internationally accepted electrical saf
9. EC tn tates 20 42cm W x 65 5cm L Gel Casting Casting sequencing gels can be achieved with the Sequencing Gel Caster or by taping around the sides and bottom of the glass plate assembly Basic laboratory safety procedures should be followed when preparing sequencing gels When working with acrylamide solutions always wear proper protective clothing gloves and eye protection Careful attention must be given during each step of sequencing gel preparation for consistent results Begin by cleaning the plates thoroughly 1 To completely clean glass plates of any acrylamide residue leftover detergents and dust wash plates with warm water and detergent Ensure plates are clean and free of any powdered detergent 2 Rinse plates completely with deionized water then wipe with 95 ethanol to eliminate any traces of detergent and water marks 3 One plate may be treated with Rain X or a siliconizing agent or Sigmacote if desired This process facilitates removal of the gel from the plates following the sequencing procedure Thermo Scientific Aluminum Sequencer 1 3 Section2 Setting Up The sequencing system arrives with the lower buffer chamber and glass separate from the upper buffer chamber and clamps To assemble follow these steps 1 2 3 4 5 6 7 8 Thermo Scientific Pull the upper piece of packing foam upwards and out of the box The upper buffer chamber assembly can then be pulled out The lower b
10. U Member State European Country and this product should be disposed of or recycled through them Further information on Thermo s compliance with this directive the recyclers in your country and information on Thermo products will be available at www thermofisher com Y Always use the proper protective equipment clothing gloves goggles etc Y Always dissipate extreme cold or heat and wear protective clothing Y Always follow good hygiene practices Y Each individual is responsible for his or her own safety Thermo Scientific Aluminum Sequencer iil Preface Do You Need Information or Assistance on Thermo Scientific Products If you do please contact us 8 00 a m to 6 00 p m Eastern Time at 1 740 373 4763 Direct 1 800 438 4851 Toll Free U S and Canada 1 877 213 8051 FAX http www thermoscientific com Internet Worldwide Web Home Page service led marietta thermofisher com Tech Support Email Address www unitylabservices com Certified Service Web Page Our Sales Support staff can provide information on pricing and give you quotations We can take your order and provide delivery information on major equipment items or make arrangements to have your local sales representative contact you Our products are listed on the Internet and we can be contacted through our Internet home page Our Service Support staff can supply technical information about proper setup operation or troubleshooting of your equipment We can fill your nee
11. ds for spare or replacement parts or provide you with on site service We can also provide you with a quotation on our Extended Warranty for your Thermo Scientific products Whatever Thermo Scientific products you need or use we will be happy to discuss your applications If you are experiencing technical problems working together we will help you locate the problem and chances are correct it yourself over the telephone without a service call When more extensive service is necessary we will assist you with direct factory trained technicians or a qualified service organization for on the spot repair If your service need is covered by the warranty we will arrange for the unit to be repaired at our expense and to your satisfaction Regardless of your needs our professional telephone technicians are available to assist you Monday through Friday from 8 00 a m to 6 00 p m Eastern Time Please contact us by telephone or fax If you wish to write our mailing address is Thermo Fisher Scientific 401 Millcreek Road Box 649 Marietta OH 45750 International customers please contact your local Thermo Scientific distributor iv Aluminum Sequencer Thermo Scientific Thermo Scientific Section 1 Section 2 Section 3 Section 4 Section 5 Section 6 Table of Contents General Information 1 1 Unpack and Check Your Order 4 2 ve SNE ER 2420 da 1 1 Gel Casting ari beg A ee a eu Tale rare 1 3 Setting Up su tween pees
12. eck the corresponding parts list Table 1 1 Maximum Buffer Volume ml Gel size W x L 35 x 45 cm 20 x 45 cm Upper chamber 450ml 300ml Thermo Scientific Aluminum Sequencer 1 1 Section 1 General Information Upper Buffer Chamber UBC Ene Well Comb 1 Power Supply Safety Lid Sharktooth Comb 2 Blank Lead Glass 2 Drain Bottle Spacer Set Safety Lid Notched Lower Buffer Glass 2 Chamber LBC Power Supply Lead Figure 1 1 Exploded Parts Diagram Model S3S Upper buffer chamber with removable drain bottle and built in side clamps Removable lower buffer chamber Safety interlocking lids with attached power supply cord 2 blank glass plates 2 notched glass plates 2 sharktooth combs 78 teeth 0 4mm thick 1 well comb 40 well 0 4mm thick 2 sets of spacers 0 4mm thick 1 adhesive temperature indicator 1 2 Aluminum Sequencer Model S4S e Upper buffer chamber with removable drain bottle and built in side clamps e Removable lower buffer chamber e Safety interlocking lids with attached power supply cord e 2 blank glass plates e 2 notched glass plates e 2 sharktooth combs 39 teeth 0 4mm thick e 1 well comb 20 well 0 4mm thick e 2 sets of spacers 0 4mm thick e 1 adhesive temperature indicator Thermo Scientific Section 1 General Information Figure 1 2 Sequencing Gel Caster Model Number 0000005 Gel Size SGE DEE 20 42cm W x 48cm L S
13. en the side clamps until the gasket is slightly flattened and a seal has been made 4 Add buffer to the upper and lower buffer chamber The buffer level in the upper buffer chamber should be high enough to submerge half the comb and fill the wells with buffer Clear any bubbles from the bottom of the gel before prerunning Using a syringe and bent needle is helpful in clearing bubbles under the gel 5 Pre run the gel Pre running serves to heat the gel and buffer to a temperature that will keep DNA in its denatured state Samples run on a gel that has not been prerun will appear distorted It is recommended that you pre run 30 to 60 minutes prior to sample loading depending on the power input A gel surface temperature of 50 60 C is considered ideal Running Conditions constant power conditions are recommended 1600 2200V 40 120mA 50 75W 1200 1600V 30 60mA 40 55W Following the pre run and before samples are loaded the gel surface must be flushed Gentle aspiration removes urea and any small bits of acrylamide that may have settled in the sample wells Loading samples may be accomplished in various ways A regular pipette tip a special flat end sequencing tip or a multichannel syringe pipette may be used The flat end sequencing tips and syringe pipettes are made to fit directly between the glass plates Care must be taken to avoid coming into contact with the gel surface when loading so the samples will enter the gel in smooth e
14. ety standard for laboratory instruments Material in this manual is for information purposes only The contents and the product it describes are subject to change without notice Thermo Fisher Scientific makes no representations or warranties with respect to this manual In no event shall Thermo be held liable for any damages direct or incidental arising out of or related to the use of this manual 2012 Thermo Fisher Scientific All rights reserved Aluminum Sequencer Thermo Scientific Preface Important operating and or maintenance instructions Read the accompanying text carefully gt gt gt Potential electrical hazards Only qualified persons should perform procedures associated with this symbol re gt Equipment being maintained or serviced must be turned off and locked off to prevent possible injury Hot surface s present which may cause burns to unprotected skin or to materials which may be damaged by elevated temperatures Marking of electrical and electronic equipment which applies to electrical and electronic equipment falling under the Directive 2002 96 EC WEEE and the equipment that has been put on the market after 13 August 2005 Lars This product is required to comply with the European Union s Waste Electrical amp Electronic Equipment WEEE Directive 2002 96 EC It is marked with the WEEE symbol Thermo Fisher Scientific has contracted with one or more recycling disposal companies in each E
15. gel polymerization Do not clamp the outer edges over the side spacers Allow the gel to completely polymerize After polymerization gently wash out the trough with buffer in order to remove any unpolymerized acrylamide and excess urea Place the comb between the glass plates in the correct left to right orientation the same orientation the trough was cast in with the comb teeth pointing down toward the gel Slowly and carefully slide the comb until it just makes contact with the gel surface without piercing it 8 Well comb Carefully insert the comb Place binder clamps over the comb and glass plates along the upper edge to force the glass plates against the comb After polymerization flood the comb with 1X TBE and remove gently Rinse the wells out with deionized water or TBE in order to remove excess urea or unpolymerized acrylamide Thermo Scientific Aluminum Sequencer 2 3 Section 2 Setting Up 2 4 Pre Running the Gel Loading Samples Aluminum Sequencer 1 After the gel has completely polymerized the gel assembly can be clamped into the sequencing unit Open up the side clamps and push them to the side Place the gel sandwich into the lower buffer chamber with the shorter offset or notched plate facing the apparatus and notched end located against UBC 2 Push the gel sandwich upright and into contact with the aluminum plate Push the side clamps toward the gel assembly and line up everything evenly 3 Screw to tight
16. king in dilute bleach DEPC is suspected to be a carcinogen and should be handled with care This electrophoresis system should never be autoclaved baked or placed in a microwave To order RNase AWAY contact Technical Services Part Number 94 e cid 04 Description ADO AAA A 250ml bottle UI 475ml spray bottle 003 uri td eer rechten beer a 1 liter bottle Les At RER REINE o aah de Ls 4 liter bottle Rnase AWAY is a registered trademark of Molecular BioProducts Aluminum Sequencer 6 3 Section 6 Care and Cleaning 6 4 Aluminum Sequencer A rn el Stopcocks 1 pair SPACER SET set includes 2 sides and 1 bottom a ws CSC Number Teeth Teeth Teeth Number Des Sharktooth 1Q_ 1 0 4 2 Sharktooth 1Q_ 1 S1S MT3 Micro Well 55 3X 0 4 Sharktooth SA 1 S1S MT44 Micro Well 73 4X 0 4 Sharktooth 10 1 S1S MT42 Micro Well 73 4X 8 and 12 channel pipette format Thermo Scientific Section 6 Optional Equipment Be EENS Stopcocks 1 pair SPACER SET set includes 2 sides and 1 bottom Se fee o Te Number Teeth Teeth Teeth Number a eG Ces Sharktooth a 1 SMIE Micro Well i ES harktooth 1 Sharktooth S 1 S25 MI44 Micro Well S harktooth Micro Well S2S MT42 8 and 12 channel pipette format Thermo Scientific Aluminum Sequencer 6 5 Section 7 Warranty Information KETEIEGED L006 OSI c1L 6 0 98 LONEULOJUI AJUCIIEM JO JOJNQIIISIP 89
17. lm exposure sometimes the bands will not be as sharp as is desired A 15 minute soak in 5 methanol and 5 acetic acid is recommended before film exposure In addition a gel run temperature that is too high can cause bands to run at different speeds causing bands to smear and drag through the gel unevenly Turning down the voltage can help eliminate this Thermo Scientific Thermo Scientific Section 5 Troubleshooting PROBLEM Curving of bands or lanes SOLUTION Curving may be caused by the uneven heating of the gel plates prior to sample loading Always preheat the gel at least 30 minutes A 50 60 C gel surface temperature should eliminate smiling and smearing For best results it is always best to avoid using the outermost lanes on the gel to eliminate any curving PROBLEM Cracking of glass SOLUTION Cracking of glass plates can be caused by overheating overtightening of clamps and leakage from the UBC Check to make sure you are running the unit within the voltages recommended When tightening clamps turning the knobs until they are just finger tight is usually enough If any sparks occur turn the power off immediately Call Technical Services PROBLEM Poor resolution SOLUTION Loading too much sample as well as using samples of poor quality may give poor gel resolution An excess of glycerol in loaded samples can pro duce a bulge effect to bands and appear as smiles or frowns in sample lanes Aluminum Seque
18. ncentrate S Aniline Benzene Butyl Acetate Calcium chloride saturated Carbon tetrachloride Chloroform Chromic acid 40 EN ES ES EM EM EE AN JEL mn E ES ES CIN ENTERAN ES pr I gt AE ECO TE EM Citric acid 10 Cottonseed oil edible This list does not include all possible chemical incompatibilities and safe compounds Acrylic products should be cleaned with warm water a mild detergent such as Alconox and can also be exposed to a mild bleach solution 10 1 In addition RNAse removal products are also safe for acrylic Contact Technical Services with any questions 6 2 Aluminum Sequencer Thermo Scientific Tips About Caring for Your System Thermo Scientific Section 6 Care and Cleaning Caution Organic solvents cause acrylic to craze or crack Clean all acrylic systems with warm water and a mild detergent Do not use ethanol or other organic solvents to clean these products Do not autoclave bake or microwave your unit Temperatures over 50 C can damage the acrylic A Note If an RNase free electrophoresis system is desired there are various methods to rid the system of RNA contamination For fast and easy decontamination use RNase AWAY Spray wipe or soak labware with RNase Away then wipe or rinse the surface clean it instantly eliminates RNase RNase Away eliminates the old methods that include treatment with 0 1 Diethyl Pyrocarbonate DEPC treated water and soa
19. ncer 5 3 Section6 Care and Cleaning Care of Acrylic The following chemical compatibility chart is supplied for the convenience of our customers Although acrylic is compatible with most solvents and solutions found in the biochemical laboratory some solvents can cause substantial damage Keep this chart handy to avoid harm to your apparatus by the use of an inappropriate solvent Codes S Safe no effect except possibly some staining A Attacked slight attack by or absorption of the liquid slight crazing or swelling but acrylic has retained most of its strength U Unsatisfactory softened swollen slowly dissolved D Dissolved in seven days or less Thermo Scientific Aluminum Sequencer 6 1 Section 6 Care and Cleaning Table 5 1 Chemical Compatibility for Acrylic Based Products Acetic acid 5 N Ethyl alcohol 95 Ethylene dichloride D Nitric acid 40 A 2 Ethylhexyl Sebacate Ss lied S Formaldehyde 40 Olive oil EN Gasoline regular leaded Phenol 5 solution D Glycerine Heptane commercial grade Hydrochloric acid 10 Hydrochloric acid concentrate Sodium hydroxide 19 Hydrogen peroxide 3 solution Sodium hydroxide 10 srw DN EN Detergent Solution Heavy Duty S Methyl alcohol 50 a Touene o we CN ION E EEN E Acetic acid Glacial Acetic Anhydride Acetone Ammonia Ammonium Chloride saturated N Ammonium Hydroxide 10 Hydroxide 10 Ammonium Hydroxide co
20. num Sequencer Notched or eared glass plates face in towards the upper buffer chamber during a gel run The ears are glass tabs on each side of the plate which prevent buffer from running out of the upper buffer chamber BLANK Offset glass plates serve the same purpose as notched plates Offset glass is usually about 2 cm shorter than the front or blank glass plate Instead of glass ears pieces of adhesive sponge are adhered either to the top of the spacers or to the apparatus itself These sponge tips wear out and may leak Notched glass plates are more fragile and NOTCHED expensive than offset glass and sponge tips but many researchers prefer notched glass because it is easier to use Figure 4 1 Glass OFFSET This is a bottom spacer and can be cut to the SPONGE TIP desired length It is optional If you use a bottom spacer when casting remember to remove it before running the gel SPACER Figure 4 2 Spacer A well comb has rectangular teeth and is similar to combs used for protein gels After polymerization these combs must be removed carefully to avoid destroying the acrylamide walls that create the wells A sharktooth comb has pointed teeth It is placed with the flat side down during gel casting to create a flat surface on the top of the gel and reinserted with the teeth down towards the gel for loading Samples are loaded in the space between the teeth Sharktooth Annnnar Standard Figure 4 3 C
21. ombs Thermo Scientific Thermo Scientific Section5 Troubleshooting Below are some possible solutions to potential problems If these suggestions are unsuccessful or you have any other questions regarding the use of this product call Technical Services PROBLEM The comb feels loose SOLUTION Try clamping across the top of the plates while the gel polymerizes This will allow the gel to polymerize more tightly around the comb These problems are usually caused by glass variability We use the highest quali ty glass available but these variations still exist Glass can also warp over time PROBLEM The gel is frowning SOLUTION There are usually two causes of this problem 1 The clamps have been overtightened or unevenly tightened causing the plates to bow Try loosening the clamps gradually making sure a seal is still present between the plates and the gasket 2 This may be caused by imperfections in the flatness of the glass plate Because of the way glass plates are made one side of the glass is usually flatter than the other Mark one side of the glass with a lab marker and try running the gel with the other side of the plate facing the acrylamide If this doesn t solve the problem the entire plate may be defective Try using a different plate PROBLEM The gel seems to be dissolving slightly at the top making it difficult to reload SOLUTION This phenomenon is probably caused by a pH imbalance Try adding fresh
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23. samples to enter the gel This serves to minimize sample diffusion and well leaking The power is then shut off and the next set is loaded until all the samples are applied Double Loading Double loading refers to the common practice of running two sets of a single sample on one sequencing gel The samples are loaded and elec trophoresis is carried out for 1 2 hours A second set of the same sample is then loaded next to the first set and electrophoresis is continued The advantage of double loading is that a greater amount of data is obtained for each sample from a single gel Sequencing Samples Sequencing samples are generally comprised of tracking dyes and glyc erol The glycerol gives density to the samples so they drop into the wells when loading The tracking dye provides color for visual monitoring of the gel run Bromophenol blue and xylene cyanol FF are the dyes used in sequencing because it has been determined that they migrate in a posi tion corresponding to DNA chain lengths On an 8 or 6 acrylamide gel the bromophenol blue migration corresponds to 20 and 30 bases respectively and is a dark blue color The xylene cyanol FF is a lighter teal blue color and its migration corresponds to 80 and 100 bases on 8 and 6 acrylamide Thermo Scientific Aluminum Sequencer 4 1 Section 4 Technical Tips The Difference between Notched and Offset Glass The Third Spacer Difference between Well 4 2 and Sharktooth Comb Alumi
24. uffer chamber is in a narrow oblong box separate from the upper buffer chamber Pull this box out also and open it up The glass is in a rectangular cardboard sheath and should be unwrapped and thoroughly cleaned before use The lower buffer chamber slides onto the base and is kept in place by four plastic stoppers The unit may be leveled using the two white nylon thumbscrews on the base and the bubble level While facing the unit gold colored plate should be facing you you will notice a small black knob approximately 1 5cm in diameter on the right side your right of the lid This knob is connected to a safety rod The purpose of which is to maintain a secure connection of the cathode to the chamber To open the lid pull the black knob upward and then slide the lid to your left You can now freely rotate the lid open In order to open the safety lid and disconnect the power to the sequencer the safety rode must be pulled up Twist open the knobs on the side clamps Push the clamps outward to allow room for your gel assembly After casting a gel the gel assembly can then be placed in the unit Aluminum Sequencer 2 1 Section 2 Setting Up Casting with Casting There are various ways to set up the glass plates and cast the sequencing Ta pe Binding Clamps gel Always make sure the comb and spacers to be used are of the same thickness Even slight variations in material may cause differences in gel thickness and cause problems
25. ven bands Regular pipette tips do not fit between the glass plates but load the sample at the opening between the two plates While resting the tip of the pipette on the shorter plate the sample is slowly dispensed The weight of the sample carries it down into the well Thermo Scientific Running the Gel Thermo Scientific Section3 Using the System 2 3 4 5 6 7 8 Run the gel as quickly as possible after loading refer to p 8 for recommended running conditions Close the upper and lower buffer chamber lids by flipping them down and sliding until the banana plug connects to the power supply lead Attach power supply leads to power supply and turn on the power When finished with the run turn off power supply and disconnect power cords Open the lids of the upper and lower buffer chambers Drain UBC into rear drain bottle by placing the hose coupling on the loose piece of hose into the stopcock attached to the male fitting on the unit This will open the port The bottle can then be removed for easy disposal Loosen the thumbscrews and slide the clamps off of the gel cassette Lift the gel cassette out of the LBC Disassemble and proceed with standard fixing drying and autoradiography The LBC may now be removed and emptied Aluminum Sequencer 3 1 Section 4 Technical Tips Sample Loading Researchers sometimes load a few sets of samples and then electrophorese them for 2 3 minutes to allow
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