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3.12 Maximum Well/Comb Volumes
NOTE: To calculate sample well volume expressed in millimeters (mm) of height, divide maximum
volume by tooth depth.
VGC-xxxx-177
# of
wells
Tooth
width
(mm)
Spacing
between
teeth (mm)
Overall
length
(mm)
Tooth
depth
(mm)
0.75mm
thickness
volume per
tooth
microliters
(ul)
1.0mm
thickness
volume per
tooth
microliters
(ul)
1.5mm
thickness
volume per
tooth
microliters
(ul)
2.0mm
thickness
volume per
tooth
microliters
(ul)
16
5.2
3.2
130
10
39
52
78
104
20
4.2
2.4
130
10
31 42 63 84
22
4.35
1.6
130
10
33 43 65 86
29
2.9
1.6
130
10
22 29 44 58
30
2.8
1.6
130
10
21 28 42 56
VGC-
XXXX
Larger Volume
Combs
# of wells
Tooth width
(mm)
Spacing
between
teeth (mm)
Overall
length
(mm)
Tooth
depth
(mm)
0.75mm
thickness
volume per
tooth
microliters (ul)
1.0mm
thickness
volume per
tooth
microliters (ul)
1.5mm
thickness
volume per
tooth
microliters (ul)
2.0mm
thickness
volume per
tooth
microliters (ul)
16
5.2
3.2
130
19.1
74
52
111
134
20
4.2
2.4
130
17.7
56 74 63 84
22
4.35
1.6
130
17.7
33 44 66 88
29
2.9
1.6
130
17.7
38 51 77 102
30
2.8
1.6
130
17.7
37 49 74 96
SECTION 4
Running Conditions
4.1 Recommended Power
1. 150V (40mA) CV for 5-7 hours
4.2 Recommended Buffers
Warning:
Do not mix buffer in the DGGE tank.
STOCK SOLUTIONS
Acrylamide Stock Solution 40% Acrylamide/Bis (37.5:1)
For 100ml use the following:
38.93 g of acrylamide
1.07 g of Bis-acrylamide
Add dH
2
O to 100ml. Do not autoclave
50x TAE Gel Running Buffer
For 1 liter use the following:
242g Tris Base
57.1ml Glacial acetic acid
100ml .5 M EDTA, pH 8.0
Add dH
2
O to 1 liter
Denaturant Stock Solution (7.5% gel)
For 100ml use:
10% 20%
30%
40%
50%
60%
70%
80%
90%
100%
40% Acrylamide/Bis (ml) 18.8
18.8
18.8
18.8
18.8
18.8
18.8
18.8
18.8
18.8
50x TAE Buffer (ml)
2
2
2
2
2
2
2
2
2
2
Formamide (ml)
4
8
12
16
20
24
28
32
36
40
Urea (g)
4.2
8.4
12.6
16.8
21 25.2 29.4 33.6 37.8
42
Add dH
2
0 to 100ml
Store denaturant stock solutions at 4º C in amber bottles
