Hoefer HE33 User Manual download pdf (Page 15)

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The electrophoresis run

Refer to the notes, buffers, and volumes section

for additional information and guidelines.



Chill the base before use, especially when higher

voltage settings will be used or when the separation

will require more than 30 minutes.

Note: To monitor separation progress, either add

0.5 µg/ml (ﬁnal conc.) of ethidium bromide to the

running buffer now, or add 50 µg/ml (ﬁnal conc.)

ethidium bromide to the sample buffer. To visualize

progress, turn off the power supply, remove the lid

assembly, and hold a portable UV lamp near the gel.

Adding ethidium bromide to the running or sample

buffer slows migration slightly. Detection by this

method is not as sensitive as staining and viewing on

a transilluminator. (See DNA detection, page 15.)



Fill both buffer chambers with running buffer until

the buffer is ~1 mm deep over the gel. (This requires

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 page 14).

Use a micro-pipette to load each sample, taking care

to avoid puncturing the well bottom or entrapping any

bubbles.



Place the lid so that the cathode (–) black lead is

at the end nearest the sample well. (Nucleic acid

samples migrate toward the anode (+) red lead.

Connect the color-coded leads (red to red, and black

to black) to an approved power supply, such as the

PS300B. Set the voltage level and timer (if available)

according to the degree of resolution sought.

Caution: Ethidium bromide is

a known mutagen. Always wear

gloves when handling.

Wear UV safety goggles and

protect skin when using a

UV lamp.

Note: If no dye was added to

the coolant, place the base on

a dark background to see the

wells more easily.

Note: At the maximum setting

the unit begins overheating

as soon as the chilled base

reaches ambient temperature.

If overheating is not controlled,

the gel will melt and/or the base

of the unit will warp!

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Hoefer HE33 User Manual Page 15

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