Nuclear Region 454 Adenita Amplification
Adenita Cultures 3 - 12
EX Takara RXN Mixture
|
conc
|
1X
|
60
|
'
|
'
|
Milli Q water |
|
4.15 |
249 |
|
|
Takara EX Buffer |
10X |
1 |
60 |
|
|
dNTP |
2.5uM |
0.8 |
48 |
|
|
Primer Mix |
20uM |
1 |
|
|
|
EX Taq |
5U |
0.05 |
3 |
|
|
Template |
Instagene |
3 |
|
|
|
|
|
|
360 |
|
|
|
|
|
30ul mix + 15ul Template |
|
|
|
|
|
9.0ul / RXN add 1.0ul Primer |
|
|
|
|
|
|
|
|
PCR cycle |
|
|
|
|
|
98C |
10 sec |
|
|
|
|
60C |
1 min |
|
|
|
|
x35 |
|
|
|
|
|
|
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|
|
|
|
|
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|
|
|
|
AMP Lane |
Template |
Primer |
AMP Lane |
Template |
Primer
|
1 |
3 Instagene |
B3 |
31 |
9 Instagene |
B9
|
2 |
3 Instagene |
C3 |
32 |
9 Instagene |
C9
|
3 |
3 Instagene |
D3 |
33 |
9 Instagene |
D9
|
4 |
3 Instagene |
E3 |
34 |
9 Instagene |
E9
|
5 |
3 Instagene |
F3 |
35 |
9 Instagene |
F9
|
6 |
4 Instagene |
B4 |
36 |
10 Instagene |
B10
|
7 |
4 Instagene |
C4 |
37 |
10 Instagene |
C10
|
8 |
4 Instagene |
D4 |
38 |
10 Instagene |
D10
|
9 |
4 Instagene |
E4 |
39 |
10 Instagene |
E10
|
10 |
4 Instagene |
F4 |
40 |
10 Instagene |
F10
|
11 |
5 Instagene |
B5 |
41 |
11 Instagene |
B11
|
12 |
5 Instagene |
C5 |
42 |
11 Instagene |
C11
|
13 |
5 Instagene |
D5 |
43 |
11 Instagene |
D11
|
14 |
5 Instagene |
E5 |
44 |
11 Instagene |
E11
|
15 |
5 Instagene |
F5 |
45 |
11 Instagene |
F11
|
16 |
6 Instagene |
B6 |
46 |
12 Instagene |
B12
|
17 |
6 Instagene |
C6 |
47 |
12 Instagene |
C12
|
18 |
6 Instagene |
D6 |
48 |
12 Instagene |
D12
|
19 |
6 Instagene |
E6 |
49 |
12 Instagene |
E12
|
20 |
6 Instagene |
F6 |
50 |
12 Instagene |
F12
|
21 |
7 Instagene |
B7 |
51 |
Neg Control |
B1
|
22 |
7 Instagene |
C7 |
52 |
Neg Control |
C1
|
23 |
7 Instagene |
D7 |
53 |
Neg Control |
D1
|
24 |
7 Instagene |
E7 |
54 |
Neg Control |
E1
|
25 |
7 Instagene |
F7 |
55 |
Neg Control |
F1
|
26 |
8 Instagene |
B8 |
56 |
Pos Control |
B1
|
27 |
8 Instagene |
C8 |
57 |
Pos Control |
C1
|
28 |
8 Instagene |
D8 |
58 |
Pos Control |
D1
|
29 |
8 Instagene |
E8 |
59 |
Pos Control |
E1
|
30 |
8 Instagene |
F8 |
60 |
Pos Control |
F1
|
|
*Positive Control = AV 20ng
|
Load 2.0ul Sample
Nuclear Region 454 Adenita Amplification
Re-AMP Adenita Culture 3 and 4
EX Takara RXN Mixture
|
conc
|
1X
|
5.5
|
Milli Q water |
|
4.15 |
22.825
|
Takara EX Buffer |
10X |
1 |
5.5
|
dNTP |
2.5uM |
0.8 |
4.4
|
Primer Mix |
20uM |
1 |
|
EX Taq |
5U |
0.05 |
0.275
|
Template |
Instagene |
3 |
16.5
|
|
|
|
49.5
|
|
|
|
9.0ul / RXN add 1.0ul Primer
|
|
|
|
|
|
|
|
|
PCR cycle |
|
|
|
98C |
10 sec |
|
|
60C |
1 min |
|
|
x35 |
|
|
|
|
|
|
|
|
|
|
|
AMP Lane |
Template |
Primer |
|
1 |
3 Instagene |
B3 |
|
2 |
3 Instagene |
C3 |
|
3 |
3 Instagene |
D3 |
|
4 |
3 Instagene |
E3 |
|
5 |
3 Instagene |
F3 |
|
6 |
4 Instagene |
B4 |
|
7 |
4 Instagene |
C4 |
|
8 |
4 Instagene |
D4 |
|
9 |
4 Instagene |
E4 |
|
10 |
4 Instagene |
F4 |
|
11 |
Neg Control |
B3 |
|
12 |
Neg Control |
C3 |
|
13 |
Neg Control |
D3 |
|
14 |
Neg Control |
E3 |
|
15 |
Neg Control |
F3 |
|
16 |
Pos Control |
B3 |
|
17 |
Pos Control |
C3 |
|
18 |
Pos Control |
D3 |
|
19 |
Pos Control |
E3 |
|
20 |
Pos Control |
F3 |
|
|
|
|
|
*Positive Control = AV 20ng |
|
|
|
Load 2.0ul Sample
Nuclear Region 454 Adenita Amplification
Re-Amp Adenita Cultures 5 & 6
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 |
|
1 |
5 Instagene |
B5 |
|
2 |
5 Instagene |
C5 |
|
3 |
5 Instagene |
D5 |
|
4 |
5 Instagene |
E5 |
|
5 |
5 Instagene |
F5 |
|
6 |
6 Instagene |
B6 |
|
7 |
6 Instagene |
C6 |
|
8 |
6 Instagene |
D6 |
|
9 |
6 Instagene |
E6 |
|
10 |
6 Instagene |
F6 |
|
11 |
Neg Control |
B5 |
|
12 |
Neg Control |
C5 |
|
13 |
Neg Control |
D5 |
|
14 |
Neg Control |
E5 |
|
15 |
Neg Control |
F5 |
|
16 |
Pos Control |
B5 |
|
17 |
Pos Control |
C5 |
|
18 |
Pos Control |
D5 |
|
19 |
Pos Control |
E5 |
|
20 |
Pos Control |
F5 |
|
|
|
|
|
*Positive Control = AV 20ng |
|
|
|
Load 2.0ul Sample
|