Nuclear Region Adenita Culture 454 Amplification
Adenita Cultures 45-48
EX Takara RXN Mixture
|
conc
|
1X
|
5
|
'
|
'
|
Milli Q water |
|
4.15 |
20.75 |
|
|
Takara EX Buffer |
10X |
1 |
5 |
|
|
dNTP |
2.5uM |
0.8 |
4 |
|
|
Primer Mix |
20uM |
1 |
|
|
|
EX Taq |
5U |
0.05 |
0.25 |
|
|
Template |
Instagene |
3 |
15 |
|
|
|
|
|
45 |
|
|
|
|
|
9.0ul / RXN add 1.0ul Primer |
|
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|
PCR cycle |
|
|
|
|
|
98C |
10 sec |
|
|
|
|
60C |
1 min |
|
|
|
|
x35 |
|
|
|
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AMP Lane |
Template |
Primer |
AMP Lane |
Template |
Primer
|
1 |
45 Instagene |
B9 |
16 |
48 Instagene |
B12
|
2 |
45 Instagene |
C9 |
17 |
48 Instagene |
C12
|
3 |
45 Instagene |
D9 |
18 |
48 Instagene |
D12
|
4 |
45 Instagene |
E9 |
19 |
48 Instagene |
E12
|
5 |
45 Instagene |
F9 |
20 |
48 Instagene |
F12
|
6 |
46 Instagene |
B10 |
21 |
Neg Control |
B9
|
7 |
46 Instagene |
C10 |
22 |
Neg Control |
C9
|
8 |
46 Instagene |
D10 |
23 |
Neg Control |
D9
|
9 |
46 Instagene |
E10 |
24 |
Neg Control |
E9
|
10 |
46 Instagene |
F10 |
25 |
Neg Control |
F9
|
11 |
47 Instagene |
B11 |
26 |
Pos Control |
B9
|
12 |
47 Instagene |
C11 |
27 |
Pos Control |
C9
|
13 |
47 Instagene |
D11 |
28 |
Pos Control |
D9
|
14 |
47 Instagene |
E11 |
29 |
Pos Control |
E9
|
15 |
47 Instagene |
F11 |
30 |
Pos Control |
F9
|
|
|
|
|
|
|
*Positive Control = AV 20ng |
|
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|
Load 2.0ul Sample
Nuclear Region Adenita Culture 454 Amplification
Re-Amp Adenita Cultures 20, 21, 33, 34, 35, 36
EX Takara RXN Mixture
|
conc
|
1X
|
5
|
'
|
'
|
Milli Q water |
|
5.15 |
25.75 |
|
|
Takara EX Buffer |
10X |
1 |
5 |
|
|
dNTP |
2.5uM |
0.8 |
4 |
|
|
Primer Mix |
20uM |
1 |
|
|
|
EX Taq |
5U |
0.05 |
0.25 |
|
|
Template |
Instagene |
2 |
10 |
|
|
|
|
|
45 |
|
|
|
|
|
9.0ul / RXN add 1.0ul Primer |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
PCR cycle |
|
|
|
|
|
98C |
10 sec |
|
|
|
|
60C |
1 min |
|
|
|
|
x35 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
AMP Lane |
Template |
Primer |
AMP Lane |
Template |
Primer
|
1 |
20 Instagene |
B8 |
21 |
35 Instagene |
B11
|
2 |
20 Instagene |
C8 |
22 |
35 Instagene |
C11
|
3 |
20 Instagene |
D8 |
23 |
35 Instagene |
D11
|
4 |
20 Instagene |
E8 |
24 |
35 Instagene |
E11
|
5 |
20 Instagene |
F8 |
25 |
35 Instagene |
F11
|
6 |
21 Instagene |
B9 |
26 |
36 Instagene |
B12
|
7 |
21 Instagene |
C9 |
27 |
36 Instagene |
C12
|
8 |
21 Instagene |
D9 |
28 |
36 Instagene |
D12
|
9 |
21 Instagene |
E9 |
29 |
36 Instagene |
E12
|
10 |
21 Instagene |
F9 |
30 |
36 Instagene |
F12
|
11 |
33 Instagene |
B9 |
31 |
Neg Control |
B8
|
12 |
33 Instagene |
C9 |
32 |
Neg Control |
C8
|
13 |
33 Instagene |
D9 |
33 |
Neg Control |
D8
|
14 |
33 Instagene |
E9 |
34 |
Neg Control |
E8
|
15 |
33 Instagene |
F9 |
35 |
Neg Control |
F8
|
16 |
34 Instagene |
B10 |
36 |
Pos Control |
B8
|
17 |
34 Instagene |
C10 |
37 |
Pos Control |
C8
|
18 |
34 Instagene |
D10 |
38 |
Pos Control |
D8
|
19 |
34 Instagene |
E10 |
39 |
Pos Control |
E8
|
20 |
34 Instagene |
F10 |
40 |
Pos Control |
F8
|
|
|
|
|
|
|
*Positive Control = AV 20ng |
|
|
|
|
|
Load 2.0ul Sample
|