Nuclear Region Adenita Culture 454 Amplification
Adenita Cultures 25-30
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 |
|
|
|
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|
45 |
|
|
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|
9.0ul / RXN add 1.0ul Primer |
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PCR cycle |
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98C |
10 sec |
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60C |
1 min |
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x35 |
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AMP Lane |
Template |
Primer |
AMP Lane |
Template |
Primer
|
1 |
25 Instagene |
B1 |
21 |
29 Instagene |
B5
|
2 |
25 Instagene |
C1 |
22 |
29 Instagene |
C5
|
3 |
25 Instagene |
D1 |
23 |
29 Instagene |
D5
|
4 |
25 Instagene |
E1 |
24 |
29 Instagene |
E5
|
5 |
25 Instagene |
F1 |
25 |
29 Instagene |
F5
|
6 |
26 Instagene |
B2 |
26 |
30 Instagene |
B6
|
7 |
26 Instagene |
C2 |
27 |
30 Instagene |
C6
|
8 |
26 Instagene |
D2 |
28 |
30 Instagene |
D6
|
9 |
26 Instagene |
E2 |
29 |
30 Instagene |
E6
|
10 |
26 Instagene |
F2 |
30 |
30 Instagene |
F6
|
11 |
27 Instagene |
B3 |
31 |
Neg Control |
B1
|
12 |
27 Instagene |
C3 |
32 |
Neg Control |
C1
|
13 |
27 Instagene |
D3 |
33 |
Neg Control |
D1
|
14 |
27 Instagene |
E3 |
34 |
Neg Control |
E1
|
15 |
27 Instagene |
F3 |
35 |
Neg Control |
F1
|
16 |
28 Instagene |
B4 |
36 |
Pos Control |
B1
|
17 |
28 Instagene |
C4 |
37 |
Pos Control |
C1
|
18 |
28 Instagene |
D4 |
38 |
Pos Control |
D1
|
19 |
28 Instagene |
E4 |
39 |
Pos Control |
E1
|
20 |
28 Instagene |
F4 |
40 |
Pos Control |
F1
|
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*Positive Control = AV 20ng |
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Load 2.0ul Sample
Nuclear Region Adenita Culture 454 Amplification
Adenita Cultures 31-36
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 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
PCR cycle |
|
|
|
|
|
98C |
10 sec |
|
|
|
|
60C |
1 min |
|
|
|
|
x35 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
AMP Lane |
Template |
Primer |
AMP Lane |
Template |
Primer
|
1 |
31 Instagene |
B7 |
21 |
35 Instagene |
B11
|
2 |
31 Instagene |
C7 |
22 |
35 Instagene |
C11
|
3 |
31 Instagene |
D7 |
23 |
35 Instagene |
D11
|
4 |
31 Instagene |
E7 |
24 |
35 Instagene |
E11
|
5 |
31 Instagene |
F7 |
25 |
35 Instagene |
F11
|
6 |
32 Instagene |
B8 |
26 |
36 Instagene |
B12
|
7 |
32 Instagene |
C8 |
27 |
36 Instagene |
C12
|
8 |
32 Instagene |
D8 |
28 |
36 Instagene |
D12
|
9 |
32 Instagene |
E8 |
29 |
36 Instagene |
E12
|
10 |
32 Instagene |
F8 |
30 |
36 Instagene |
F12
|
11 |
33 Instagene |
B9 |
31 |
Neg Control |
B7
|
12 |
33 Instagene |
C9 |
32 |
Neg Control |
C7
|
13 |
33 Instagene |
D9 |
33 |
Neg Control |
D7
|
14 |
33 Instagene |
E9 |
34 |
Neg Control |
E7
|
15 |
33 Instagene |
F9 |
35 |
Neg Control |
F7
|
16 |
34 Instagene |
B10 |
36 |
Pos Control |
B7
|
17 |
34 Instagene |
C10 |
37 |
Pos Control |
C7
|
18 |
34 Instagene |
D10 |
38 |
Pos Control |
D7
|
19 |
34 Instagene |
E10 |
39 |
Pos Control |
E7
|
20 |
34 Instagene |
F10 |
40 |
Pos Control |
F7
|
|
|
|
|
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|
*Positive Control = AV 20ng |
|
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Load 2.0ul Sample
Nuclear Region Adenita Culture 454 Amplification
Re-Amp Adenita Cultures 11, 17, 18 ,19 with 2.0ul Instagene Template
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 |
11 Instagene |
B11 |
16 |
19 Instagene |
B7
|
2 |
11 Instagene |
C11 |
17 |
19 Instagene |
C7
|
3 |
11 Instagene |
D11 |
18 |
19 Instagene |
D7
|
4 |
11 Instagene |
E11 |
19 |
19 Instagene |
E7
|
5 |
11 Instagene |
F11 |
20 |
19 Instagene |
F7
|
6 |
17 Instagene |
B5 |
21 |
Neg Control |
B11
|
7 |
17 Instagene |
C5 |
22 |
Neg Control |
C11
|
8 |
17 Instagene |
D5 |
23 |
Neg Control |
D11
|
9 |
17 Instagene |
E5 |
24 |
Neg Control |
E11
|
10 |
17 Instagene |
F5 |
25 |
Neg Control |
F11
|
11 |
18 Instagene |
B6 |
26 |
Pos Control |
B11
|
12 |
18 Instagene |
C6 |
27 |
Pos Control |
C11
|
13 |
18 Instagene |
D6 |
28 |
Pos Control |
D11
|
14 |
18 Instagene |
E6 |
29 |
Pos Control |
E11
|
15 |
18 Instagene |
F6 |
30 |
Pos Control |
F11
|
|
|
|
|
|
|
*Positive Control = AV 20ng |
|
|
|
|
|
Load 2.0ul Sample
|