2 recommended buffers – C.B.S. Scientific MGU-602T User Manual
Page 10
www.cbsscientific.com
Tel# 858-755-4959
9
3.4 Maximum Well/Comb Volumes
NOTE: To calculate sample well volume expressed in millimeters (mm) of height, divide maximum
volume by tooth depth.
MBU
Comb for
MGU-
402T
# of wells
Tooth
Depth
mm
Overall
length
of comb
mm
Spacing
between
teeth
mm
Tooth
width/
mm
1.0mm
thickness
volume per
tooth
microliters
(ul)
1.5mm
thickness
volume
per tooth
microliters
(ul)
2.0mm
thickness
volume per
tooth
microliters
(ul)
3.0mm
thickness
volume per
tooth
microliters
(ul)
5
12.7
87.28
1.58
16.2
205.74
308.61
411.48
617.22
12
12.7
87.28
1.58
5.83
74.04
111.06
148.08
222.12
18
12.7
87.28
1.58
3.37
42.79
64.19
85.58
128.37
SGC14
Comb for
MGU-
502T &
MGU-
602T
# of wells
Tooth
Depth
mm
Overall
length
of comb
mm
Spacing
between
teeth
mm
Tooth
width/
mm
1.0mm
thickness
volume per
tooth
microliters
(ul)
1.5mm
thickness
volume
per tooth
microliters
(ul)
2.0mm
thickness
volume per
tooth
microliters
(ul)
3.0mm
thickness
volume per
tooth
microliters
(ul)
1
12.7
120.64
3
120
1524
2286
3048
4672
3E
12.7
120.64
3
38.1
483.87
725.81
967.74
1451.61
5
12.7
120.64
3
21.6
274
411.48
548
822
10
12.7
120.64
3
9.2
117
175.26
232
350
12
12.7
120.64
2.4
7.8
99
148.59
198
297
13
12.7
120.64
2.4
6.6
83.82
125.73
167.64
251.46
16
12.7
120.64
2.4
5.3
67.3
100.9
134.6
201
20
12.7
120.64
1.6
4.5
57
85.7
114
171
26
12.7
120.64
1.6
3.1
39.3
59
78
118
27
12.7
120.64
1.6
2.93
37.21
55.82
74.42
111.63
SECTION 4
Running Conditions
4.1 Recommended Power
As a rule, optimal resolution of larger molecules is achieved during longer runs at lower voltages,
whereas smaller molecules require shorter runs at higher voltages. Applied voltage gradients can
therefore be anywhere in the range of 1-10 VDC/cm of gel. Using the standard buffer systems
listed below, most runs will use 5 VDC/cm as a general rule.
The usual run time will vary for the voltage chosen but should range from 15 to 60 minutes.
Nucleic acid migration is monitored by the progress of marker dyes. Constant power is not a
necessity, but it produces uniform heat throughout the run, therefore minimizing band diffusion.
Be sure the polarity is correct i.e. that the DNA is loaded near the cathode (black electrode) to run
toward the anode (red terminal).
Agarose gels may be stored for several days at 4°C wrapped in plastic wrap. Seakem
Agarose (FMC) is used (normally) for preparative and analytical gels. Other types of
agarose can be used for special purposes.
4.2
Recommended Buffers*
Type*
Concentrated Stock/liter
Final Concentration
TAE
50X - 242 gm Tris base
1X - 0.04M Tris-acetate
(Tris-acetate) 57.1ml glacial acetic acid
0.001M EDTA
100 ml 0.5M EDTA (pH 8.0)
TBE
5X - 54 gm Tris base
0.5X - 0.045M Tris-borate
(Tris-borate)
27.5 gm boric acid
0.001M EDTA
20 ml 0.5M EDTA (pH 8.0)
10X - Loading Buffer (DNA)*
0.25% Bromophenol blue
0.25% Xylene cyanol
20% Ficoll Type 400
0.1M EDTA, pH 8.0