User:Alexander S Mikheyev/Notebook/rotifer alien genes/2010/05/11

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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
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

11May10.jpg


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

11May10B.jpg

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

11May10C.jpg