...
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | ||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
A | GBS10-11 PCR_A Col_1 | GBS10-11 PCR_A Col_6 | GBS10-11 PCR_A Col_12 | GBS10-11 PCR_B Col_1 | GBS10-11 PCR_B Col_6 | GBS10-11 PCR_B Col_12 |
| GBS1-4 PCR_A Col_1 | GBS1-4 PCR_B Col_1 |
| NTC | NTC | NTC | |
B | GBS10-11 PCR_A Col_1 | GBS10-11 PCR_A Col_6 | GBS10-11 PCR_A Col_12 | GBS10-11 PCR_B Col_1 | GBS10-11 PCR_B Col_6 | GBS10-11 PCR_B Col_12 |
| GBS1-4 PCR_A Col_1 | GBS1-4 PCR_B Col_1 |
| 0.0002 pM Std | 0.0002 pM Std | 0.0002 pM Std | |
C | GBS10-11 PCR_A Col_1 | GBS10-11 PCR_A Col_6 | GBS10-11 PCR_A Col_12 | GBS10-11 PCR_B Col_1 | GBS10-11 PCR_B Col_6 | GBS10-11 PCR_B Col_12 |
| GBS1-4 PCR_A Col_1 | GBS1-4 PCR_B Col_1 |
| 0.002 pM Std | 0.002 pM Std | 0.002 pM Std | |
D | GBS10-11 PCR_A Col_1 | GBS10-11 PCR_A Col_6 | GBS10-11 PCR_A Col_12 | GBS10-11 PCR_B Col_1 | GBS10-11 PCR_B Col_6 | GBS10-11 PCR_B Col_12 |
|
| GBS1-4 PCR_A Col_1 | GBS1-4 PCR_B Col_1 |
| 0.02 pM Std | 0.02 pM Std | 0.02 pM Std |
E | GBS10-11 PCR_A Col_1 | GBS10-11 PCR_A Col_6 | GBS10-11 PCR_A Col_12 | GBS10-11 PCR_B Col_1 | GBS10-11 PCR_B Col_6 | GBS10-11 PCR_B Col_12 |
| GBS1-4 PCR_A Col_1 | GBS1-4 PCR_B Col_1 |
| 0.2 pM Std | 0.2 pM Std | 0.2 pM Std | |
F | GBS10-11 PCR_A Col_1 | GBS10-11 PCR_A Col_6 | GBS10-11 PCR_A Col_12 | GBS10-11 PCR_B Col_1 | GBS10-11 PCR_B Col_6 | GBS10-11 PCR_B Col_12 |
|
| GBS1-4 PCR_A Col_1 | GBS1-4 PCR_B Col_1 |
| 2 pM Std | 2 pM Std | 2 pM Std |
G | GBS10-11 PCR_A Col_1 | GBS10-11 PCR_A Col_6 | GBS10-11 PCR_A Col_12 | GBS10-11 PCR_B Col_1 | GBS10-11 PCR_B Col_6 | GBS10-11 PCR_B Col_12 |
|
| GBS1-4 PCR_A Col_1 | GBS1-4 PCR_B Col_1 |
| 20 pM Std | 20 pM Std | 20 pM Std |
H | GBS10-11 PCR_A Col_1 | GBS10-11 PCR_A Col_6 | GBS10-11 PCR_A Col_12 | GBS10-11 PCR_B Col_1 | GBS10-11 PCR_B Col_6 | GBS10-11 PCR_B Col_12 |
|
| GBS1-4 PCR_A Col_1 | GBS1-4 PCR_B Col_1 |
|
|
|
|
Combine 5uL from each well of GBS10-11_PCR_A into a pool. Make a 1:1000 dilution of this pool to run on qPCR.
Combine 5uL from each well of GBS10-11_PCR_B into a pool. Make a 1:1000 dilution of this pool to run on qPCR.
After 1:1000 pools have been made for each PCR plate, combine PCR_A and PCR_B undiluted pools together into one tube and label GBS10-11. Make a 1:1000 dilution of this pool to run on qPCR.
Add 16 ul of Illumina Library Quantification MasterMix to each well:
...
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | ||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
A | GBS10-11 PCR_A Col_1 | GBS10-11 PCR_A Col_6 | GBS10-11 PCR_A Col_12 | GBS10-11 PCR_B Col_1 | GBS10-11 PCR_B Col_6 | GBS10-11 PCR_B Col_12 | GBS10-11 PCR_A Pool |
|
| GBS1-4 PCR_A Col_1 | GBS1-4 PCR_B Col_1 | NTC | NTC | NTC |
B | GBS10-11 PCR_A Col_1 | GBS10-11 PCR_A Col_6 | GBS10-11 PCR_A Col_12 | GBS10-11 PCR_B Col_1 | GBS10-11 PCR_B Col_6 | GBS10-11 PCR_B Col_12 | GBS10-11 PCR_A Pool |
| GBS1-4 PCR_A Col_1 | GBS1-4 PCR_B Col_1 | 0.0002 pM Std | 0.0002 pM Std | 0.0002 pM Std | |
C | GBS10-11 PCR_A Col_1 | GBS10-11 PCR_A Col_6 | GBS10-11 PCR_A Col_12 | GBS10-11 PCR_B Col_1 | GBS10-11 PCR_B Col_6 | GBS10-11 PCR_B Col_12 | GBS10-11 PCR_B Pool |
| GBS1-4 PCR_A Col_1 | GBS1-4 PCR_B Col_1 | 0.002 pM Std | 0.002 pM Std | 0.002 pM Std | |
D | GBS10-11 PCR_A Col_1 | GBS10-11 PCR_A Col_6 | GBS10-11 PCR_A Col_12 | GBS10-11 PCR_B Col_1 | GBS10-11 PCR_B Col_6 | GBS10-11 PCR_B Col_12 | GBS10-11 PCR_B Pool |
|
| GBS1-4 PCR_A Col_1 | GBS1-4 PCR_B Col_1 | 0.02 pM Std | 0.02 pM Std | 0.02 pM Std |
E | GBS10-11 PCR_A Col_1 | GBS10-11 PCR_A Col_6 | GBS10-11 PCR_A Col_12 | GBS10-11 PCR_B Col_1 | GBS10-11 PCR_B Col_6 | GBS10-11 PCR_B Col_12 | GBS10-11 Pool |
| GBS1-4 PCR_A Col_1 | GBS1-4 PCR_B Col_1 | 0.2 pM Std | 0.2 pM Std | 0.2 pM Std | |
F | GBS10-11 PCR_A Col_1 | GBS10-11 PCR_A Col_6 | GBS10-11 PCR_A Col_12 | GBS10-11 PCR_B Col_1 | GBS10-11 PCR_B Col_6 | GBS10-11 PCR_B Col_12 | GBS10-11 Pool |
|
| GBS1-4 PCR_A Col_1 | GBS1-4 PCR_B Col_1 | 2 pM Std | 2 pM Std | 2 pM Std |
G | GBS10-11 PCR_A Col_1 | GBS10-11 PCR_A Col_6 | GBS10-11 PCR_A Col_12 | GBS10-11 PCR_B Col_1 | GBS10-11 PCR_B Col_6 | GBS10-11 PCR_B Col_12 |
|
|
| GBS1-4 PCR_A Col_1 | GBS1-4 PCR_B Col_1 | 20 pM Std | 20 pM Std | 20 pM Std |
H | GBS10-11 PCR_A Col_1 | GBS10-11 PCR_A Col_6 | GBS10-11 PCR_A Col_12 | GBS10-11 PCR_B Col_1 | GBS10-11 PCR_B Col_6 | GBS10-11 PCR_B Col_12 |
|
|
| GBS1-4 PCR_A Col_1 | GBS1-4 PCR_B Col_1 |
|
|
|
Results:
Average quantities observed for all products:
...
GBS1-4 PCR_A Col_1:
GBS1-4 PCR_B Col_1:
...
Bioanalyzer
BioAnalyzer Chip Set up:
Well 1: GBS10-11 PCR_A Pool
...