Batch 3 Sample Information
Sample order in plate | Sample No. | [RNA] (ng/ul) | Vol RNA used | Vol H2O to add | ng RNA used |
---|---|---|---|---|---|
1 | 35 | 94.4 | 3.71 | 1.29 | 350 |
2 | 524 | 63.0 | 5.00 | – | 315 |
3 | 453 | 196.0 | 1.79 | 3.21 | 350 |
4 | 554 | 188.0 | 1.86 | 3.14 | 350 |
5 | 442b | 69.8 | 5.00 | – | 349 |
6 | 489 | 68.0 | 5.00 | – | 340 |
7 | 462b | 106.0 | 3.30 | 1.70 | 350 |
8 | 533 | 6.5 | 5.00 | – | 32.6 |
9 | 522 | 32.2 | 5.00 | – | 161 |
10 | 474 | 77.2 | 4.53 | 0.47 | 350 |
11 | 452b | 97.2 | 3.60 | 1.40 | 350 |
12 | 443 | 60.2 | 5.00 | – | 301 |
13 | 477 | 164.0 | 2.13 | 2.87 | 350 |
14 | 526 | 138.0 | 2.54 | 2.46 | 350 |
15 | 562 | 126.0 | 2.78 | 2.22 | 350 |
16 | 432 | 74.0 | 4.73 | 0.27 | 350 |
17 | 37 | 74.6 | 4.69 | 0.31 | 350 |
18 | 413 | 130.0 | 2.69 | 2.31 | 350 |
19 | 45 | 25.1 | 5.00 | – | 125.5 |
20 | 561 | 28.0 | 5.00 | – | 140 |
21 | 542 | 32.8 | 5.00 | – | 164 |
22 | 527 | 124.0 | 2.82 | 2.18 | 350 |
23 | 492 | 82.6 | 4.24 | 0.76 | 350 |
24 | 475 | 27.4 | 5.00 | – | 137 |
25 | 541 | 44.4 | 5.00 | – | 222 |
26 | 565 | 31.2 | 5.00 | – | 156 |
27 | B3 – NTC1 | NA | – | 5.00 | #VALUE! |
28 | B3 – NTC2 | NA | – | 5.00 | #VALUE! |
Batch 3 Plate Configuration
1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 |
---|---|---|---|---|---|---|---|
A | 35 | 524 | 453 | 554 | 442b | 489 | 462b |
B | |||||||
C | 533 | 522 | 474 | 452b | 443 | 477 | 526 |
D | |||||||
E | 562 | 432 | 37 | 413 | 45 | 561 | 542 |
F | |||||||
G | 527 | 492 | 475 | 541 | 565 | B3 – NTC1 | B3 – NTC2 |
H |
Batch 4 Sample Information
Sample order in plate | Sample No. | [RNA] (ng/ul) | Vol RNA used | Vol H2O to add | ng RNA used |
---|---|---|---|---|---|
1 | 571 | LOW | 5.00 | – | NA |
2 | 525 | 140.0 | 2.50 | 2.50 | 350 |
3 | 563 | 47.0 | 5.00 | – | 235 |
4 | 404 | 112.0 | 3.13 | 1.88 | 350 |
5 | 484 | 58.4 | 5.00 | – | 292 |
6 | 531 | 95.4 | 3.67 | 1.33 | 350 |
7 | 34 | 104.0 | 3.37 | 1.63 | 350 |
8 | 490 | 186.0 | 1.88 | 3.12 | 350 |
9 | 523 | 71.4 | 4.90 | 0.10 | 350 |
10 | 473 | 124.0 | 2.82 | 2.18 | 350 |
11 | 485 | 19.1 | 5.00 | – | 95.5 |
12 | 402 | 114.0 | 3.07 | 1.93 | 350 |
13 | 487 | 118.0 | 2.97 | 2.03 | 350 |
14 | 476 | 39.2 | 5.00 | – | 196 |
15 | 421 | 57.6 | 5.00 | – | 288 |
16 | 553 | 186.0 | 1.88 | 3.12 | 350 |
17 | 41 | 108.0 | 3.24 | 1.76 | 350 |
18 | 46 | 98.6 | 3.55 | 1.45 | 350 |
19 | 551 | 96.4 | 3.63 | 1.37 | 350 |
20 | 486b | 148.0 | 2.36 | 2.64 | 350 |
21 | 445 | 160.0 | 2.19 | 2.81 | 350 |
22 | 43 | 156.0 | 2.24 | 2.76 | 350 |
23 | 506 | 29.2 | 5.00 | – | 146 |
24 | 482 | 22.2 | 5.00 | – | 111 |
25 | 412 | 31.2 | 5.00 | – | 156 |
26 | 488 | 60.0 | 5.00 | – | 300 |
27 | 47 | 128.0 | 2.73 | 2.27 | 350 |
28 | B4 – NTC2 | NA | – | 5.00 | 0 |
Batch 4 Plate Configuration
1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 |
---|---|---|---|---|---|---|---|
A | 571 | 525 | 563 | 404 | 484 | 531 | 34 |
B | |||||||
C | 490 | 523 | 473 | 485 | 402 | 487 | 476 |
D | |||||||
E | 421 | 553 | 41 | 46 | 551 | 486b | 445 |
F | |||||||
G | 43 | 506 | 482 | 412 | 488 | 47 | B4 – NTC2 |
H |
Notes
- Aliquoted 20 uL RNA into new tubes and froze in -80. Except, for the following samples I aliquoted 7 uL: 34, 35, 41, 46, 46
- Sample #35 was not fully sealed during either the RNA removal step or the second strand synthesis step, and I believe I lost some volume to evaporation. I proceeded normally, but will see how that library looks.
- Not sure if I mentioned this previously, but I’m using 350 ng RNA, except for low concentration samples (I use 5 uL to get the maximum amount of RNA possible).
- To “quickly spin down” samples at room temperature I put the sample plate in the centrifuge, push start, then push stop immediately when the speed does not read 0. I also set the temperature to room temperature so it doesn’t actively cool.
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