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E3xxx

E3xxx

99

Solutions

Solutions

1x TAE  40 mM tris (pH 7.6), 20 mM acetic acid, 1 mM EDTA

• 50 x (1l) dissolve in 750 ml distilled water:
• 242 g tris base (FW = 121)
• 57.1 ml glacial acetic acid
• 100 ml 0.5 M EDTA (pH 8.0).
• Fill to 1 litre with distilled water.

1x TBE 89 mM tris (pH 7.6), 89 mM boric acid, 2 mM EDTA

• 10 x (1l) dissolve in 750 ml distilled water:
• 108 g tris base (FW = 121)
• 55 g boric acid (FW = 61.8)
• 40 ml 0.5 M EDTA (pH 8.0)
• Fill to 1 litre with distilled water.

Sample Loading Dye

• 10x  sample  buffer  stock  consists  of  50%  glycerol,  0.25% 

bromophenol  blue,  and  0.25%  xylene  cyanole  FF  in  1x  TAE 

buffer.  Only  1–10  ml  of  the  10x  loading  dye  should  be  pre-

pared.

Ethidium Bromide Solution

• Add 10 mg of Ethidium Bromide to 1 ml distilled water.

Содержание E3100

Страница 1: ...Consort MANUAL E3100 E3200 E3300 E3400 E3500 E3600 July 2009 Distributor Topac Inc 231 CJC Highway Cohasset MA 02025 Tel 781 740 8778 Fax 781 740 8779 www topac com email sales topac com ...

Страница 2: ...tal conditions 3 Fitting electrode cables 4 Fitting loading guides 4 Gel preparation 5 Gel pouring with casting dams 6 Gel pouring with model E3100 6 Gel pouring using tape method 7 Gel pouring using the flexicaster 7 Running the gel 8 Gel staining and viewing 8 Solutions 9 Troubleshooting 10 ...

Страница 3: ...d maintenance procedures out lined in this instruction manual and when not having been subject to accident alteration misuse or abuse No liability is accepted for loss or damage arising from the incor rect use of this unit CONSORT s liability is limited to the repair or replacement of the unit or refund of the purchase price at CONSORT s option CONSORT is not liable for any consequential damages C...

Страница 4: ...connect the leads Do not exceed the maximum operating voltage or current Do not operate the electrophoresis units in metal trays Acrylamide is a volatile cumulative neurotoxin and suspect ed carcinogen Wear effective protective clothing and follow recommended handling and disposal procedures Polymerised gels contain some unpolymerised monomer Han dle with gloves only Following the replacement of a...

Страница 5: ...re should be taken not to damage the enclosed electrode and vigorous cleaning is not necessary or advised Air drying is preferably before use The units should only be cleaned with the following warm water with a mild concentration of soap or other mild de tergent compatible detergents include dish washing liquid hexane and aliphatic hydrocarbons The units should not be left to in detergents for mo...

Страница 6: ...p between the lid and the leading edge of no gap between the lid and the leading edge of the cable fitting 4 Refit the lid Fitting loading guides Fitting loading guides These can be fitted to enhance visibility of the wells if desired They can be fitted to the white vinyl platform sheet or to the unit itself 1 Seat the tray in the unit and note the position of the comb grooves The samples run blac...

Страница 7: ...ical flask 3 Add the appropriate amount of 1 x TAE or TBE solution from the table above To prevent evaporation during the dissolving steps below the conical flask should be covered with para film 4 Dissolve the agarose powder by heating the agarose either on a magnetic hot plate with stirring bar or in a microwave oven If using the microwave method the microwave should be set at around a 400 W or ...

Страница 8: ...tank the gel including tray to the main tank Gel pouring Gel pouring with model E3100 with model E3100 Gel pouring with model E3100 Gel pouring Gel pouring with model E3100 Gel pouring 1 Fit the casting gates into the grooves in the unit which are 10 mm from the platinum wire Ensure that these are seated as deeply as possible This will ensure that a good seal is formed and that there is no possibi...

Страница 9: ...agarose to set ensuring that the gel remains undis turbed 5 Carefully remove the gel casting gates and comb and transfer the gel including tray to the main tank the gel including tray to the main tank Gel pouring using the flexicaster Gel pouring using the flexicaster 1 Level the flexicaster base by adjusting the feet so that the bubble is exactly central 2 Insert the desired length tray into the ...

Страница 10: ... faint then the staining pro cedure should be adjusted so that there is less de staining If there is too much background then the staining procedure should be adjusted so that there is more de staining Running the gel Running the gel 1 Mix the sample to be loaded with sample buffer see solutions for common sample buffers Usually 3 µl of sample buffer is adequate but less may be used with sample vo...

Страница 11: ... tris pH 7 6 89 mM boric acid 2 mM EDTA 10 x 1l dissolve in 750 ml distilled water 108 g tris base FW 121 55 g boric acid FW 61 8 40 ml 0 5 M EDTA pH 8 0 Fill to 1 litre with distilled water Sample Loading Dye 10x sample buffer stock consists of 50 glycerol 0 25 bromophenol blue and 0 25 xylene cyanole FF in 1x TAE buffer Only 1 10 ml of the 10x loading dye should be pre pared Ethidium Bromide Sol...

Страница 12: ...nt gel from breaking Avoid damaging the well with the pipette when loading the sample Aim for the centre of the well and avoid damaging the bottom of the well with the pipette tip Samples leak underneath the gel upon loading The bottom of the wells were torn when the comb was re moved To avoid this tearing carefully wiggle the comb to free the teeth from the gel Distorted sample wells Incomplete p...

Страница 13: ...erol in the sample Poor band resolution Add ficoll glycerol or sucrose to the sample loading buffer to ensure that the sample layers on the bottom of the well Ensure sample is completely dissolved Reduce voltage sample concentration or sample volume Ensure there is at least 1 mm of gel below the bottom of the comb to prevent samples from leaking out the bottom of the well Reduce salt concentration...

Страница 14: ...12 E3xxx E3xxx ...

Страница 15: ... E3100 E3200 E3300 E3400 E3500 E3600 to which this declaration relates is in conformity with the following standards EN61010 LOW VOLTAGE DIRECTIVE 73 23 EEG EN50081 1 EN50082 1 EN60555 2 EMC DIRECTIVE 89 336 EEG Turnhout June 25 2009 CONSORT nv Tel 32 14 41 12 79 Parklaan 36 Fax 32 14 42 91 79 B 2300 Turnhout E mail info consort be ...

Страница 16: ......

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