User:Alexander S Mikheyev/Notebook/rotifer alien genes/2010/05/17

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Nuclear Region Adenita Culture 454 Amplification

Adenita Cultures 45-48

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 45 Instagene B9 16 48 Instagene B12
2 45 Instagene C9 17 48 Instagene C12
3 45 Instagene D9 18 48 Instagene D12
4 45 Instagene E9 19 48 Instagene E12
5 45 Instagene F9 20 48 Instagene F12
6 46 Instagene B10 21 Neg Control B9
7 46 Instagene C10 22 Neg Control C9
8 46 Instagene D10 23 Neg Control D9
9 46 Instagene E10 24 Neg Control E9
10 46 Instagene F10 25 Neg Control F9
11 47 Instagene B11 26 Pos Control B9
12 47 Instagene C11 27 Pos Control C9
13 47 Instagene D11 28 Pos Control D9
14 47 Instagene E11 29 Pos Control E9
15 47 Instagene F11 30 Pos Control F9
*Positive Control = AV 20ng

Load 2.0ul Sample

17May10A.jpg

Nuclear Region Adenita Culture 454 Amplification

Re-Amp Adenita Cultures 20, 21, 33, 34, 35, 36

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 20 Instagene B8 21 35 Instagene B11
2 20 Instagene C8 22 35 Instagene C11
3 20 Instagene D8 23 35 Instagene D11
4 20 Instagene E8 24 35 Instagene E11
5 20 Instagene F8 25 35 Instagene F11
6 21 Instagene B9 26 36 Instagene B12
7 21 Instagene C9 27 36 Instagene C12
8 21 Instagene D9 28 36 Instagene D12
9 21 Instagene E9 29 36 Instagene E12
10 21 Instagene F9 30 36 Instagene F12
11 33 Instagene B9 31 Neg Control B8
12 33 Instagene C9 32 Neg Control C8
13 33 Instagene D9 33 Neg Control D8
14 33 Instagene E9 34 Neg Control E8
15 33 Instagene F9 35 Neg Control F8
16 34 Instagene B10 36 Pos Control B8
17 34 Instagene C10 37 Pos Control C8
18 34 Instagene D10 38 Pos Control D8
19 34 Instagene E10 39 Pos Control E8
20 34 Instagene F10 40 Pos Control F8
*Positive Control = AV 20ng

Load 2.0ul Sample

17May10B.jpg