Sample loading buffer, Loading buffer, Table 3: comb specifications and well volumes – Hoefer HE33 User Manual
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Sample loading buffer
Loading buffer
(5X, 25% Ficoll 400, 0.25% Bromophenol 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 to sample in proportion so that 1/5 of the final
volume is loading buffer. (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 and dragging
effects caused by comigration with smaller nucleic acids.
Table 3: Comb specifications and well volumes
well
sample vol.
comb
no. of
thickness width
per 1 mm
code no.
wells
(mm)
(mm)
depth (µl)
HE31A-P-1.0 1 prep/2 ref 1.0
44/6
44/6*
HE31A-P-1.5 1 prep/2 ref 1.5
44/6
66/9*
HE31A-8-1.0 8
1.0
6.5
6.5
HE31A-8-1.5 8
1.5
6.5
9.7
HE31A-12-1.0 12
1.0
3.9
3.9
HE31A-12-1.5 12
1.5
3.9
5.8
HE31A-16-1.0 16
1.0
2.6
2.6
HE31A-16-1.5 16
1.5
2.6
3.9
* The preparative combs form two reference wells (for MW
standards), one on each side of the preparative well. The
first number is sample volume/mm in the preparative well;
the second is volume/mm in the reference well.