Nuclear Gene Rotifer Culture 454 Amplification
Adenita Culture 13-24
| EX Takara RXN Mixture
|
conc
|
1X
|
6.5
|
'
|
| Milli Q water |
|
5.15 |
33.475 |
|
| Takara EX Buffer |
10X |
1 |
6.5 |
|
| dNTP |
2.5uM |
0.8 |
5.2 |
|
| Primer Mix |
20uM |
1 |
|
|
| EX Taq |
5U |
0.05 |
0.325 |
|
| Template |
Instagene |
2 |
13 |
|
|
|
|
58.5 |
9.0ul / RXN
|
| A PCR cycle |
|
|
|
|
| 98C |
10 sec |
|
|
|
| 60C |
1 min |
|
|
|
| x35 |
|
|
|
|
|
|
|
|
|
| Plate 2 |
|
|
|
|
| Source Plate |
Well |
EX ID |
|
|
| Ithaca 1 |
A2 |
13 |
|
|
| Ithaca 1 |
A3 |
14 |
|
|
| Ithaca 1 |
A5 |
15 |
|
|
| Ithaca 1 |
A6 |
16 |
|
|
| Ithaca 1 |
B1 |
17 |
|
|
| Ithaca 1 |
B3 |
18 |
|
|
| Ithaca 1 |
B5 |
19 |
|
|
| Ithaca 1 |
C1 |
20 |
|
|
| Ithaca 1 |
D1 |
21 |
|
|
| Ithaca 1 |
D3 |
22 |
|
|
| Ithaca 1 |
D4 |
23 |
|
|
| Ithaca 1 |
D6 |
24 |
|
|
Shearing by NEB fragmentase
| Reaction mixture
|
conc.
|
1x (ul)
|
x4
|
| DNA (2ug) |
363 ng/ul |
5.5 |
22
|
| Fragmentase buffer |
10x |
2 |
8
|
| BSA |
100x |
0.2 |
0.8
|
| MilliQ |
|
10.3 |
41.2
|
|
|
|
on ice |
5 min |
|
|
|
| Fragmentase |
|
2 |
|
|
|
|
37C |
No. 1 |
10 min
|
|
|
No. 2 |
15 min
|
|
|
No. 3 |
20 min
|
|
|
No. 4 |
25 min
|
File:2100 expert High Sensitivity DNA Assay DE72901887 2010-04-28.pdf
|
Alien genes
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