User:Alexander S Mikheyev/Notebook/rotifer alien genes/2010/05/12

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Nuclear Gene 454 Adenita Amplification

Re-Amp Cultures 7-10

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 7 Instagene B7
2 7 Instagene C7
3 7 Instagene D7
4 7 Instagene E7
5 7 Instagene F7
6 8 Instagene B8
7 8 Instagene C8
8 8 Instagene D8
9 8 Instagene E8
10 8 Instagene F8
11 9 Instagene B9
12 9 Instagene C9
13 9 Instagene D9
14 9 Instagene E9
15 9 Instagene F9
16 10 Instagene B10
17 10 Instagene C10
18 10 Instagene D10
19 10 Instagene E10
20 10 Instagene F10
21 Neg Control B7
22 Neg Control C7
23 Neg Control D7
24 Neg Control E7
25 Neg Control F7
26 Pos Control B7
27 Pos Control C7
28 Pos Control D7
29 Pos Control E7
30 Pos Control F7
*Positive Control = AV 20ng


Load 2.0ul Sample

12May10.jpg

Nuclear Gene 454 Adenita Amplification

Re-Amp Cultures 11-12

Amp Cultures 13-16

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 11 Instagene B11
2 11 Instagene C11
3 11 Instagene D11
4 11 Instagene E11
5 11 Instagene F11
6 12 Instagene B12
7 12 Instagene C12
8 12 Instagene D12
9 12 Instagene E12
10 12 Instagene F12
11 13 Instagene B1
12 13 Instagene C1
13 13 Instagene D1
14 13 Instagene E1
15 13 Instagene F1
16 14 Instagene B2
17 14 Instagene C2
18 14 Instagene D2
19 14 Instagene E2
20 14 Instagene F2
21 15 Instagene B3
22 15 Instagene C3
23 15 Instagene D3
24 15 Instagene E3
25 15 Instagene F3
26 16 Instagene B4
27 16 Instagene C4
28 16 Instagene D4
29 16 Instagene E4
30 16 Instagene F4
31 Neg Control B1
32 Neg Control C1
33 Neg Control D1
34 Neg Control E1
35 Neg Control F1
36 Pos Control B1
37 Pos Control C1
38 Pos Control D1
39 Pos Control E1
40 Pos Control F1
*Positive Control = AV 20ng

Load 2.0ul Sample

12May10B.jpg

Nuclear Gene 454 Adenita Amplification

Amp Cultures 17-24

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 17 Instagene B5 26 22 Instagene B10
2 17 Instagene C5 27 22 Instagene C10
3 17 Instagene D5 28 22 Instagene D10
4 17 Instagene E5 29 22 Instagene E10
5 17 Instagene F5 30 22 Instagene F10
6 18 Instagene B6 31 23 Instagene B11
7 18 Instagene C6 32 23 Instagene C11
8 18 Instagene D6 33 23 Instagene D11
9 18 Instagene E6 34 23 Instagene E11
10 18 Instagene F6 35 23 Instagene F11
11 19 Instagene B7 36 24 Instagene B12
12 19 Instagene C7 37 24 Instagene C12
13 19 Instagene D7 38 24 Instagene D12
14 19 Instagene E7 39 24 Instagene E12
15 19 Instagene F7 40 24 Instagene F12
16 20 Instagene B8 41 Neg Control B5
17 20 Instagene C8 42 Neg Control C5
18 20 Instagene D8 43 Neg Control D5
19 20 Instagene E8 44 Neg Control E5
20 20 Instagene F8 45 Neg Control F5
21 21 Instagene B9 46 Pos Control B5
22 21 Instagene C9 47 Pos Control C5
23 21 Instagene D9 48 Pos Control D5
24 21 Instagene E9 49 Pos Control E5
25 21 Instagene F9 50 Pos Control F5
*Positive Control = AV 20ng

Load 2.0ul Sample

12May10C.jpg