Nuclear Gene Adenita Culture 454 Amplification
Re-Amp Instagene Cultures 57, C, D, I
EX Takara RXN Mixture
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conc
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1X
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5
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'
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'
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Milli Q water |
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4.15 |
20.75 |
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Takara EX Buffer |
10X |
1 |
5 |
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dNTP |
2.5uM |
0.8 |
4 |
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Primer Mix |
20uM |
1 |
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EX Taq |
5U |
0.05 |
0.25 |
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Template |
Instagene |
3 |
15 |
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45 |
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9.0ul / RXN add 1.0ul Primer |
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PCR cycle |
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98C |
10 sec |
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60C |
1 min |
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x35 |
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AMP Lane |
Template |
Primer |
AMP Lane |
Template |
Primer
|
1 |
Instagene 57 |
B3 |
16 |
Instagene I |
B3
|
2 |
Instagene 57 |
C3 |
17 |
Instagene I |
C3
|
3 |
Instagene 57 |
D3 |
18 |
Instagene I |
D3
|
4 |
Instagene 57 |
E3 |
19 |
Instagene I |
E3
|
5 |
Instagene 57 |
F3 |
20 |
Instagene I |
F3
|
6 |
Instagene C |
B3 |
26 |
Neg Control |
B3
|
7 |
Instagene C |
C3 |
27 |
Neg Control |
C3
|
8 |
Instagene C |
D3 |
28 |
Neg Control |
D3
|
9 |
Instagene C |
E3 |
29 |
Neg Control |
E3
|
10 |
Instagene C |
F3 |
30 |
Neg Control |
F3
|
11 |
Instagene D |
B3 |
31 |
Pos Control |
B3
|
12 |
Instagene D |
C3 |
32 |
Pos Control |
C3
|
13 |
Instagene D |
D3 |
33 |
Pos Control |
D3
|
14 |
Instagene D |
E3 |
34 |
Pos Control |
E3
|
15 |
Instagene D |
F3 |
35 |
Pos Control |
F3
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*Positive Control = AV 20ng |
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Load 2.0ul Sample
Nuclear Gene Adenita Culture 454 Amplification QIAquick Cleaning
1) Combined products from primer 2-6 for each culture
2) Cleaned with Qiagen QIAquick PCR product purification kit
http://www.qiagen.com/products/dnacleanup/gelpcrsicleanupsystems/qiaquickpcrpurificationkit.aspx#Tabs=t2
see pg 19
Nanodrop results
Sample ID
|
Nucleic Acid Conc.
|
Unit
|
A260
|
A280
|
260/280
|
260/230
|
Sample Type
|
Factor
|
1 |
75.2 |
ng/µl |
1.504 |
0.823 |
1.83 |
2.15 |
DNA |
50
|
2 |
60.5 |
ng/µl |
1.211 |
0.665 |
1.82 |
2.17 |
DNA |
50
|
3 |
90.6 |
ng/µl |
1.812 |
1.014 |
1.79 |
2.2 |
DNA |
50
|
4 |
97.2 |
ng/µl |
1.945 |
1.071 |
1.82 |
2.18 |
DNA |
50
|
5 |
52.8 |
ng/µl |
1.055 |
0.584 |
1.81 |
2.14 |
DNA |
50
|
6 |
82.6 |
ng/µl |
1.653 |
0.892 |
1.85 |
2.07 |
DNA |
50
|
7 |
75.3 |
ng/µl |
1.506 |
0.813 |
1.85 |
2.08 |
DNA |
50
|
8 |
56.5 |
ng/µl |
1.129 |
0.634 |
1.78 |
2.15 |
DNA |
50
|
9 |
87.5 |
ng/µl |
1.75 |
0.953 |
1.84 |
2.23 |
DNA |
50
|
10 |
65.2 |
ng/µl |
1.305 |
0.725 |
1.8 |
2.09 |
DNA |
50
|
11 |
51.5 |
ng/µl |
1.029 |
0.574 |
1.79 |
1.87 |
DNA |
50
|
12 |
44.4 |
ng/µl |
0.889 |
0.488 |
1.82 |
2.14 |
DNA |
50
|
13 |
51.9 |
ng/µl |
1.039 |
0.573 |
1.81 |
1.93 |
DNA |
50
|
14 |
54.9 |
ng/µl |
1.098 |
0.6 |
1.83 |
2.19 |
DNA |
50
|
15 |
63.8 |
ng/µl |
1.277 |
0.703 |
1.82 |
2.21 |
DNA |
50
|
16 |
55.1 |
ng/µl |
1.102 |
0.594 |
1.86 |
2.27 |
DNA |
50
|
17 |
64.6 |
ng/µl |
1.292 |
0.7 |
1.85 |
2.25 |
DNA |
50
|
18 |
68.3 |
ng/µl |
1.366 |
0.741 |
1.84 |
2.19 |
DNA |
50
|
19 |
79.1 |
ng/µl |
1.583 |
0.867 |
1.83 |
2.18 |
DNA |
50
|
20 |
58.6 |
ng/µl |
1.173 |
0.653 |
1.8 |
2.14 |
DNA |
50
|
21 |
45.1 |
ng/µl |
0.902 |
0.498 |
1.81 |
1.84 |
DNA |
50
|
22 |
66.2 |
ng/µl |
1.323 |
0.734 |
1.8 |
2.2 |
DNA |
50
|
23 |
70 |
ng/µl |
1.4 |
0.755 |
1.86 |
2.19 |
DNA |
50
|
24 |
45.3 |
ng/µl |
0.906 |
0.499 |
1.82 |
2.16 |
DNA |
50
|
25 |
44.3 |
ng/µl |
0.887 |
0.487 |
1.82 |
2.21 |
DNA |
50
|
26 |
66.2 |
ng/µl |
1.323 |
0.722 |
1.83 |
2.14 |
DNA |
50
|
27 |
49.8 |
ng/µl |
0.995 |
0.545 |
1.83 |
2.16 |
DNA |
50
|
28 |
51.7 |
ng/µl |
1.034 |
0.567 |
1.82 |
2.05 |
DNA |
50
|
29 |
45.2 |
ng/µl |
0.905 |
0.506 |
1.79 |
1.96 |
DNA |
50
|
30 |
76.8 |
ng/µl |
1.535 |
0.837 |
1.83 |
2.11 |
DNA |
50
|
31 |
48.4 |
ng/µl |
0.968 |
0.544 |
1.78 |
2.12 |
DNA |
50
|
32 |
37.3 |
ng/µl |
0.746 |
0.419 |
1.78 |
1.96 |
DNA |
50
|
33 |
28.7 |
ng/µl |
0.574 |
0.329 |
1.75 |
1.72 |
DNA |
50
|
34 |
38.5 |
ng/µl |
0.771 |
0.428 |
1.8 |
1.93 |
DNA |
50
|
35 |
39.5 |
ng/µl |
0.789 |
0.446 |
1.77 |
1.85 |
DNA |
50
|
36 |
44.2 |
ng/µl |
0.883 |
0.479 |
1.85 |
2.13 |
DNA |
50
|
37 |
42.6 |
ng/µl |
0.853 |
0.471 |
1.81 |
2.07 |
DNA |
50
|
38 |
32.3 |
ng/µl |
0.646 |
0.357 |
1.81 |
1.92 |
DNA |
50
|
39 |
49.1 |
ng/µl |
0.982 |
0.546 |
1.8 |
1.99 |
DNA |
50
|
40 |
43.7 |
ng/µl |
0.875 |
0.483 |
1.81 |
1.98 |
DNA |
50
|
41 |
48.6 |
ng/µl |
0.971 |
0.548 |
1.77 |
2.08 |
DNA |
50
|
42 |
46.2 |
ng/µl |
0.925 |
0.502 |
1.84 |
2.15 |
DNA |
50
|
43 |
43.5 |
ng/µl |
0.87 |
0.49 |
1.78 |
1.96 |
DNA |
50
|
44 |
46.7 |
ng/µl |
0.934 |
0.516 |
1.81 |
2.12 |
DNA |
50
|
45 |
55.1 |
ng/µl |
1.103 |
0.591 |
1.87 |
1.98 |
DNA |
50
|
46 |
52.5 |
ng/µl |
1.05 |
0.584 |
1.8 |
2.12 |
DNA |
50
|
47 |
37.6 |
ng/µl |
0.751 |
0.42 |
1.79 |
2.02 |
DNA |
50
|
48 |
50.1 |
ng/µl |
1.002 |
0.548 |
1.83 |
2.07 |
DNA |
50
|
49 |
59.7 |
ng/µl |
1.194 |
0.652 |
1.83 |
2.11 |
DNA |
50
|
50 |
33.8 |
ng/µl |
0.676 |
0.385 |
1.75 |
1.97 |
DNA |
50
|
51 |
42.4 |
ng/µl |
0.848 |
0.481 |
1.76 |
1.93 |
DNA |
50
|
52 |
66 |
ng/µl |
1.321 |
0.729 |
1.81 |
2.16 |
DNA |
50
|
53 |
81.2 |
ng/µl |
1.625 |
0.895 |
1.81 |
2.06 |
DNA |
50
|
54 |
42.1 |
ng/µl |
0.842 |
0.476 |
1.77 |
2.05 |
DNA |
50
|
55 |
39 |
ng/µl |
0.78 |
0.458 |
1.7 |
1.8 |
DNA |
50
|
56 |
28.9 |
ng/µl |
0.579 |
0.347 |
1.67 |
1.91 |
DNA |
50
|
57 |
51.6 |
ng/µl |
1.031 |
0.577 |
1.79 |
2.12 |
DNA |
50
|
58 |
52.3 |
ng/µl |
1.045 |
0.602 |
1.74 |
2.12 |
DNA |
50
|
59 |
57.5 |
ng/µl |
1.15 |
0.639 |
1.8 |
2.1 |
DNA |
50
|
60 |
57.2 |
ng/µl |
1.144 |
0.644 |
1.78 |
2.06 |
DNA |
50
|
61 |
49.4 |
ng/µl |
0.989 |
0.577 |
1.71 |
2.03 |
DNA |
50
|
62 |
59.8 |
ng/µl |
1.196 |
0.674 |
1.78 |
1.91 |
DNA |
50
|
63 |
42.7 |
ng/µl |
0.855 |
0.491 |
1.74 |
1.99 |
DNA |
50
|
64 |
45.5 |
ng/µl |
0.91 |
0.517 |
1.76 |
1.8 |
DNA |
50
|
65 |
43.8 |
ng/µl |
0.877 |
0.49 |
1.79 |
1.91 |
DNA |
50
|
66 |
37.4 |
ng/µl |
0.748 |
0.419 |
1.79 |
2.07 |
DNA |
50
|
67 |
37.9 |
ng/µl |
0.757 |
0.437 |
1.74 |
1.86 |
DNA |
50
|
68 |
31 |
ng/µl |
0.619 |
0.368 |
1.68 |
1.8 |
DNA |
50
|
69 |
39.4 |
ng/µl |
0.789 |
0.445 |
1.77 |
2.05 |
DNA |
50
|
70 |
44.8 |
ng/µl |
0.895 |
0.521 |
1.72 |
1.63 |
DNA |
50
|
71 |
37.8 |
ng/µl |
0.755 |
0.444 |
1.7 |
1.86 |
DNA |
50
|
72 |
41.8 |
ng/µl |
0.836 |
0.481 |
1.74 |
2.03 |
DNA |
50
|
73 |
64.2 |
ng/µl |
1.284 |
0.72 |
1.78 |
2.06 |
DNA |
50
|
74 |
40.1 |
ng/µl |
0.803 |
0.452 |
1.78 |
1.82 |
DNA |
50
|
75 |
33.6 |
ng/µl |
0.673 |
0.382 |
1.76 |
2.06 |
DNA |
50
|
76 |
26.5 |
ng/µl |
0.531 |
0.31 |
1.71 |
2 |
DNA |
50
|
A |
51.5 |
ng/µl |
1.029 |
0.571 |
1.8 |
2.17 |
DNA |
50
|
B |
51.6 |
ng/µl |
1.033 |
0.575 |
1.8 |
1.86 |
DNA |
50
|
C |
79.3 |
ng/µl |
1.587 |
0.889 |
1.79 |
2.22 |
DNA |
50
|
D |
55.2 |
ng/µl |
1.105 |
0.626 |
1.77 |
2.01 |
DNA |
50
|
E |
49 |
ng/µl |
0.98 |
0.561 |
1.75 |
2.05 |
DNA |
50
|
F |
36.7 |
ng/µl |
0.735 |
0.438 |
1.68 |
1.83 |
DNA |
50
|
G |
51.5 |
ng/µl |
1.029 |
0.59 |
1.74 |
2.04 |
DNA |
50
|
H |
48.7 |
ng/µl |
0.974 |
0.56 |
1.74 |
1.88 |
DNA |
50
|
I |
61.2 |
ng/µl |
1.224 |
0.695 |
1.76 |
2.05 |
DNA |
50
|
CCA3 |
72.9 |
ng/µl |
1.458 |
0.807 |
1.81 |
2.11 |
DNA |
50
|
A. ricciae |
105.2 |
ng/µl |
2.104 |
1.147 |
1.84 |
2.21 |
DNA |
50
|
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