Loading buffer and sample volumes – Hoefer HE99X User Manual
Page 20
Loading buffer and sample volumes
Loading buffer
(5X, 25% Ficoll 400, 0.25% Bromphenol blue
†
, 10 ml)
Deionized H
2
O
to 7.0 ml
Ficoll 400
2.5 g
Bromophenol blue (FW 691.9)
25.0 mg
Deionized H
2
O
to 10.0 ml
Add 1 volume loading buffer to 4 volumes of sample.
(Loading buffer increases solution density.)
Note 1: Sucrose or glycerol may be used instead of
Ficoll 400.
Note 2: Xylene cyanol (0.25%), which migrates
more slowly than bromophenol blue, can be
added as an additional marker if desired. The
agarose concentration determines the position
of the dye bands relative to a polynucleotide.
†
Tracking dyes may be omitted to eliminate obscuring or dragging
effects caused by comigration with smaller nucleic acids.
Comb specifications and well volumes
well
well
sample vol.
code
no. of
width
thickness per 1 mm
number
wells
(mm)
(mm)
depth (µl)
HE91A-P-1.5
1/2
113/10
1.5
171/14.5*
HE91A-P-3.0
1/2
113/10
3.0
342/29.0*
HE91A-10-1.5
10
9.7
1.5
14.5
HE91A-10-3.0
10
9.7
3.0
30.0
HE91A-15-1.0
15
7.1
1.0
7.1
HE91A-15-1.5
15
7.1
1.5
10.6
HE91A-15-3.0
15
7.1
3.0
21.3
HE91A-20-1.0
20
4.7
1.0
4.7
HE91A-20-1.5
20
4.7
1.5
7.1
HE91A-20-3.0
20
4.7
3.0
14.2
HE91A-30-1.0
30
3.0
1.0
3.0
* Preparative combs form two reference wells (for MW standards), one on
each side of the preparative well. The first number is sample volume/
mm depth in the preparative well; the second is volume/mm in the
reference well.
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