User:Alexander S Mikheyev/Notebook/rotifer alien genes/2010/05/13

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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
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 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
*Positive Control = AV 20ng

Load 2.0ul Sample

13May10.jpg

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
*Positive Control = AV 20ng


Load 2.0ul Sample

13May10B.jpg

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

13May10C.jpg