Hoefer HE33 User Manual Download

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user

manual

nucleic acid electrophoresis

HE33-IM/Rev. H0/05-04

Hoefer HE 33

mini submarine electrophoresis unit

Summary of Contents for HE33

Page 1: ...user manual nucleic acid electrophoresis um HE33 IM Rev H0 05 04 Hoefer HE 33 mini submarine electrophoresis unit...

Page 2: ......

Page 3: ...ne electrophoresis unit function 1 Important information 2 Unpacking 4 Specifications 4 Operating instructions 5 Care and maintenance 11 Troubleshooting 12 Notes buffers and volumes 13 Bibliography 17...

Page 4: ...pii...

Page 5: ...ion The Hoefer HE 33 horizontal agarose unit is intended for rapid electrophoresis of small quantities of nucleic acids in agarose gels A gel is cast in the gel caster which holds one or two combs Eig...

Page 6: ...in the base it may crack Organic solvents will cause irreparable chemical damage to the unit Do not chill the base below 20 C 4 F Chill the base in a bucket of ice refrigerator or in a freezer Do not...

Page 7: ...solvants organiques peuvent causer des dommages chimiques irr parables Ne pas refroidir le socle en dessous de 20 C 4 F Refroidir le socle dans un seau rempli de glace ou un dans un r frig rateur Ne p...

Page 8: ...500 mA Max operating temp 50 C Max buffer volume 250 ml Coolant required 600 ml 50 50 water ethylene glycol Gel size 7 10 cm Environmental operating conditions Indoor use 4 40 C Humidity up to 80 Alti...

Page 9: ...ary heat sink Prepare 600 ml of 50 50 ethylene glycol water Optional To help see wells more clearly while loading the sample add a drop or two of soluble dye or food color to the coolant solution Loca...

Page 10: ...ow the solution to cool to 50 C before pouring into the casting tray Optional Add 0 5 g ml ethidium bromide to the gel solution to observe separation during electrophoresis Caution Ethidium bromide is...

Page 11: ...tray Tighten the screws to secure the comb To run twice as many samples prepare two combs Remove the comb assembly Place the casting assembly on a leveling surface and level using the spirit level on...

Page 12: ...t Transfer the running tray and gel to the chilled base comb back comb screws 2 Fig 3 A comb back which fits onto the rim of the casting tray positions the comb in the gel Two screws hold the adjustab...

Page 13: ...ires about 220 ml Load the samples Add sample to 5X sample loading buffer and mix 1 5 of the final volume is loading buffer see p 14 Use a micro pipette to load each sample taking care to avoid punctu...

Page 14: ...n 20 to 30 minutes Chill the base before use Table 1 Voltage settings and recommended run settings voltage gradient time V V cm min 500 40 5 400 31 10 300 24 20 200 16 30 to 40 150 12 30 to 60 For rap...

Page 15: ...gen peroxide H2O2 solution then rinse thoroughly with DEPC treated autoclaved deionized water Sambrook et al 1 7 40 Replacing foam pads Remove worn foam pads Peel off the adhesive cover on a new foam...

Page 16: ...the casting tray before pouring the gel Double banded pattern The comb must be vertical to prevent well shape distortion Decrease the buffer level to 1 mm above the top of the gel to reduce the verti...

Page 17: ...olt age setting temperature conformation and the presence of ethidium bromide Special agaroses are available that can extend resolu tion ranges A common standard is a Hind III digest of lambda phage w...

Page 18: ...for DNA in agarose gels Recipes for the two most commonly used running buffers for DNA electrophoresis are listed below The ionic strength of these buffers is appropriate for the application do not ad...

Page 19: ...0 5 M pH 8 0 100 ml Na2EDTA 2H2O FW 372 2 0 5 M 18 6 g Deionized H2O to 70 0 ml NaOH 10 M to pH 8 0 5 0 ml Deionized H2O to 100 0 ml Sample loading buffer Loading buffer 5X 25 Ficoll 400 0 25 Bromophe...

Page 20: ...eparative well The first number is sample volume mm in the preparative well the second is volume mm in the reference well DNA detection DNA can be detected either by the fluorescence of bound ethidium...

Page 21: ...d intensity 302 nm UV light to reduce photonicking To reduce the background fluorescence of unbound ethidium bromide the gel can be destained by soaking it for 5 minutes in 0 01 m MgCl2 or for 1 hour...

Page 22: ...one 8 well 1 5 mm thick comb comb back and screws Replacement parts Buffer chamber assembly HE30 Lid with high voltage leads HE36 Fill plug bottom HE38 Fill plug top HE38TP Gel running tray UVT 7 10...

Page 23: ...P 1 0 1 0 8 HE31A 8 1 0 1 0 12 HE31A 12 1 0 1 0 16 HE31A 16 1 0 1 5 Preparative HE31A P 1 5 1 5 8 HE31A 8 1 5 1 5 12 HE31A 12 1 5 1 5 16 HE31A 16 1 5 Comb back with 2 screws HE31 BK Replacement screw...

Page 24: ...Hoefer Inc 953 Indiana Street San Francisco CA 94107 USA www hoeferinc com Printed in the USA...