User:Alexander S Mikheyev/Notebook/rotifer alien genes/2010/05/14

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Nuclear Region Adenita Culture 454 Amplification

Adenita Cultures 37-40

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 37 Instagene B1 16 40 Instagene B4
2 37 Instagene C1 17 40 Instagene C4
3 37 Instagene D1 18 40 Instagene D4
4 37 Instagene E1 19 40 Instagene E4
5 37 Instagene F1 20 40 Instagene F4
6 38 Instagene B2 21 Neg Control B1
7 38 Instagene C2 22 Neg Control C1
8 38 Instagene D2 23 Neg Control D1
9 38 Instagene E2 24 Neg Control E1
10 38 Instagene F2 25 Neg Control F1
11 39 Instagene B3 26 Pos Control B1
12 39 Instagene C3 27 Pos Control C1
13 39 Instagene D3 28 Pos Control D1
14 39 Instagene E3 29 Pos Control E1
15 39 Instagene F3 30 Pos Control F1
*Positive Control = AV 20ng

Load 2.0ul Sample

14May10A.jpg

Nuclear Region Adenita Culture 454 Amplification

Re-Amp 2.0ul Adenita Cultures 22-25

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 22 Instagene B10 16 25 Instagene B1
2 22 Instagene C10 17 25 Instagene C1
3 22 Instagene D10 18 25 Instagene D1
4 22 Instagene E10 19 25 Instagene E1
5 22 Instagene F10 20 25 Instagene F1
6 23 Instagene B11 21 Neg Control B1
7 23 Instagene C11 22 Neg Control C1
8 23 Instagene D11 23 Neg Control D1
9 23 Instagene E11 24 Neg Control E1
10 23 Instagene F11 25 Neg Control F1
11 24 Instagene B12 26 Pos Control B1
12 24 Instagene C12 27 Pos Control C1
13 24 Instagene D12 28 Pos Control D1
14 24 Instagene E12 29 Pos Control E1
15 24 Instagene F12 30 Pos Control F1
*Positive Control = AV 20ng

Load 2.0ul Sample

14May10B.jpg

Nuclear Region Adenita Culture 454 Amplification

Re-Amp 2.0ul Adenita Cultures 27-32

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 27 Instagene B3 21 31 Instagene B7
2 27 Instagene C3 22 31 Instagene C7
3 27 Instagene D3 23 31 Instagene D7
4 27 Instagene E3 24 31 Instagene E7
5 27 Instagene F3 25 31 Instagene F7
6 28 Instagene B4 26 32 Instagene B8
7 28 Instagene C4 27 32 Instagene C8
8 28 Instagene D4 28 32 Instagene D8
9 28 Instagene E4 29 32 Instagene E8
10 28 Instagene F4 30 32 Instagene F8
11 29 Instagene B5 31 Neg Control B3
12 29 Instagene C5 32 Neg Control C3
13 29 Instagene D5 33 Neg Control D3
14 29 Instagene E5 34 Neg Control E3
15 29 Instagene F5 35 Neg Control F3
16 30 Instagene B6 36 Pos Control B3
17 30 Instagene C6 37 Pos Control C3
18 30 Instagene D6 38 Pos Control D3
19 30 Instagene E6 39 Pos Control E3
20 30 Instagene F6 40 Pos Control F3
*Positive Control = AV 20ng

Load 2.0ul Sample

14May10C.jpg