...
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 |
|---|---|---|---|---|---|---|---|---|---|---|---|---|
A | ALF_PLT5_TN_Col116S_Col3 | ALF_PLT5_TN_Col516S_Col9 | ALF_PLT5_TN_ITS_Col7Col3 | ALF_PLT5_TN_Col11ITS_Col9 | ALF_PLT12_TN_16S_Col1Col3 | ALF_PLT12_TN_Col516S_Col9 | ALF_PLT12_TN_ITS_Col7Col3 | ALF_PLT12_TN_Col11ITS_Col9 |
|
|
| NTC |
B | ALF_PLT5_TN_Col116S_Col3 | ALF_PLT5_TN_16S_Col5Col9 | ALF_PLT5_TN_Col7ITS_Col3 | ALF_PLT5_TN_ITS_Col11Col9 | ALF_PLT12_TN_Col116S_Col3 | ALF_PLT12_TN_16S_Col5Col9 | ALF_PLT12_TN_Col7ITS_Col3 | ALF_PLT12_TN_ITS_Col11Col9 |
|
| 04.0002 pM Std | 0.0002 pM Std |
C | ALF_PLT5_TN_Col116S_Col3 | ALF_PLT5_TN_Col516S_Col9 | ALF_PLT5_TN_Col7ITS_Col3 | ALF_PLT5_TN_Col11ITS_Col9 | ALF_PLT12_TN_16S_Col1Col3 | ALF_PLT12_TN_Col516S_Col9 | ALF_PLT12_TN_ITS_Col7Col3 | ALF_PLT12_TN_Col11ITS_Col9 |
|
| 0.002 pM Std | 0.002 pM Std |
D | ALF_PLT5_TN_16S_Col1Col3 | ALF_PLT5_TN_Col516S_Col9 | ALF_PLT5_TN_Col7ITS_Col3 | ALF_PLT5_TN_Col11ITS_Col9 | ALF_PLT12_TN_Col116S_Col3 | ALF_PLT12_TN_16S_Col5Col9 | ALF_PLT12_TN_Col7ITS_Col3 | ALF_PLT12_TN_ITS_Col11Col9 |
|
| 0.02 pM Std | 0.02 pM Std |
E | ALF_PLT5_TN_Col116S_Col3 | ALF_PLT5_TN_16S_Col5Col9 | ALF_PLT5_TN_ITS_Col7Col3 | ALF_PLT5_TN_ITS_Col11Col9 | ALF_PLT12_TN_Col116S_Col3 | ALF_PLT12_TN_Col516S_Col9 | ALF_PLT12_TN_ITS_Col7Col3 | ALF_PLT12_TN_Col11ITS_Col9 |
|
| 0.2 pM Std | 0.2 pM Std |
F | ALF_PLT5_TN_16S_Col1Col3 | ALF_PLT5_TN_Col516S_Col9 | ALF_PLT5_TN_ITS_Col7Col3 | ALF_PLT5_TN_Col11ITS_Col9 | ALF_PLT12_TN_16S_Col1Col3 | ALF_PLT12_TN_Col516S_Col9 | ALF_PLT12_TN_Col7ITS_Col3 | ALF_PLT12_TN_ITS_Col11Col9 |
|
| 2 pM Std | 2 pM Std |
G | ALF_PLT5_TN_Col116S_Col3 | ALF_PLT5_TN_Col516S_Col9 | ALF_PLT5_TN_Col7ITS_Col3 | ALF_PLT5_TN_ITS_Col11Col9 | ALF_PLT12_TN_16S_Col1Col3 | ALF_PLT12_TN_16S_Col5Col9 | ALF_PLT12_TN_Col7ITS_Col3 | ALF_PLT12_TN_ITS_Col11Col9 |
|
| 20 pM Std | 20 pM Std |
H | ALF_PLT5_TN_Col116S_Col3 | ALF_PLT5_TN_Col516S_Col9 | ALF_PLT5_TN_ITS_Col7Col3 | ALF_PLT5_TN_Col11ITS_Col9 | ALF_PLT12_TN_16S_Col1Col3 | ALF_PLT12_TN_Col516S_Col9 | ALF_PLT12_TN_ITS_Col7Col3 | ALF_PLT12_TN_Col11ITS_Col9 |
|
|
|
|
Add 16 ul of Illumina Library Quantification MasterMix to each well:
...
Add 4 ul of template, pool, or standards to each well:
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 |
|---|
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 |
|---|---|---|---|---|---|---|---|---|---|---|---|---|
A | ALF_PLT5_TN_16S_Col1Col3 | ALF_PLT5_TN_Col516S_Col9 | ALF_PLT5_TN_Col7ITS_Col3 | ALF_PLT5_TN_ITS_Col11Col9 | ALF_PLT12_TN_Col116S_Col3 | ALF_PLT12_TN_16S_Col5Col9 | ALF_PLT12_TN_Col7ITS_Col3 | ALF_PLT12_TN_ITS_Col11Col9 |
| NTC | NTC | NTC |
B | ALF_PLT5_TN_Col116S_Col3 | ALF_PLT5_TN_16S_Col5Col9 | ALF_PLT5_TN_Col7ITS_Col3 | ALF_PLT5_TN_Col11ITS_Col9 | ALF_PLT12_TN_16S_Col1Col3 | ALF_PLT12_TN_Col516S_Col9 | ALF_PLT12_TN_ITS_Col7Col3 | ALF_PLT12_TN_Col11ITS_Col9 |
| 0.0002 pM Std | 0.0002 pM Std | 0.0002 pM Std |
C | ALF_PLT5_TN_16S_Col1Col3 | ALF_PLT5_TN_Col516S_Col9 | ALF_PLT5_TN_ITS_Col7Col3 | ALF_PLT5_TN_Col11ITS_Col9 | ALF_PLT12_TN_Col116S_Col3 | ALF_PLT12_TN_16S_Col5Col9 | ALF_PLT12_TN_Col7ITS_Col3 | ALF_PLT12_TN_ITS_Col11Col9 |
| 0.002 pM Std | 0.002 pM Std | 0.002 pM Std |
D | ALF_PLT5_TN_Col116S_Col3 | ALF_PLT5_TN_16S_Col5Col9 | ALF_PLT5_TN_Col7ITS_Col3 | ALF_PLT5_TN_ITS_Col11Col9 | ALF_PLT12_TN_Col116S_Col3 | ALF_PLT12_TN_Col516S_Col9 | ALF_PLT12_TN_ITS_Col7Col3 | ALF_PLT12_TN_Col11ITS_Col9 |
| 0.02 pM Std | 0.02 pM Std | 0.02 pM Std |
E | ALF_PLT5_TN_Col116S_Col3 | ALF_PLT5_TN_Col516S_Col9 | ALF_PLT5_TN_ITS_Col7Col3 | ALF_PLT5_TN_ITS_Col11Col9 | ALF_PLT12_TN_16S_Col1Col3 | ALF_PLT12_TN_Col516S_Col9 | ALF_PLT12_TN_ITS_Col7Col3 | ALF_PLT12_TN_Col11ITS_Col9 |
| 0.2 pM Std | 0.2 pM Std | 0.2 pM Std |
F | ALF_PLT5_TN_Col116S_Col3 | ALF_PLT5_TN_16S_Col5Col9 | ALF_PLT5_TN_Col7ITS_Col3 | ALF_PLT5_TN_ITS_Col11Col9 | ALF_PLT12_TN_Col116S_Col3 | ALF_PLT12_TN_16S_Col5Col9 | ALF_PLT12_TN_Col7ITS_Col3 | ALF_PLT12_TN_ITS_Col11Col9 |
| 2 pM Std | 2 pM Std | 2 pM Std |
G | ALF_PLT5_TN_Col116S_Col3 | ALF_PLT5_TN_Col516S_Col9 | ALF_PLT5_TN_Col7ITS_Col3 | ALF_PLT5_TN_Col11ITS_Col9 | ALF_PLT12_TN_Col116S_Col3 | ALF_PLT12_TN_16S_Col5Col9 | ALF_PLT12_TN_ITS_Col7Col3 | ALF_PLT12_TN_Col11ITS_Col9 |
| 20 pM Std | 20 pM Std | 20 pM Std |
H | ALF_PLT5_TN_16S_Col1Col3 | ALF_PLT5_TN_Col516S_Col9 | ALF_PLT5_TN_ITS_Col7Col3 | ALF_PLT5_TN_Col11ITS_Col9 | ALF_PLT12_TN_Col116S_Col3 | ALF_PLT12_TN_16S_Col5Col9 | ALF_PLT12_TN_Col7ITS_Col3 | ALF_PLT12_TN_Col11ITS_Col9 |
|
|
|
|
Results:
Average results are below:
ALF_PLT5_TN_Col116S_Col3:
ALF_PLT5_TN_16S_Col5Col9:
ALF_PLT5_TN_Col7ITS_Col3:
ALF_PLT5_TN_ITS_Col11Col9:
ALF_PLT12_TN_Col116S_Col3:
ALF_PLT12_TN_Col516S_Col9:
ALF_PLT12_TN_Col7ITS_Col3:
ALF_PLT12_TN_ITS_Col11Col9:
iSeq Run:
Dilute to 1 nM based off qPCR results. qPCR results are in pM, but 1:1000 dilution used. The results are effectively in nM for pool.
...