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ul/rxn | Reagent | # of rxns | ul needed | |
|---|---|---|---|---|
3 | 5X Kapa HiFi Buffer | 2200 | 6600 | 2640 |
0.45 | 10M dNTPs | 2200 | 990396 | |
0.3 | Kapa HiFi HotStart DNA Pol | 2200 | 660 | 264 |
5.25 | HPLC H2O | 2200 | 159507260 | |
9 | Total Volume | 2200 | 24200 | 10,560 |
Add 9 ul to each well of a hard shell, full skirt plate. Seal with bubble strips and store in refrigerator until needed label “NS5 PCR”. (If doing manually, one needs 2 ul of template and 2 ul of the primers).
Plate | 16S Primer | ITS Primer |
|---|---|---|
HW1 | 16S0A1 | ITS0A1 |
16S0B1 | ITS0B1 | |
HW2 | 16S0C1 | ITS0C1 |
16S0D1 | ITS0D1 | |
NW1 | 16S0E1 | ITS0E1 |
16S0F1 | ITS0F1 | |
NW2 | 16S0G1 | ITS0G1 |
16S0H1 | ITS0H1 | |
NW3 | 16S0A2 | ITS0A2 |
16S0B2 | ITS0B2 | |
NW4 | 16S0C2 | ITS0C2 |
16S0D2 | ITS0D2 | |
BW1 | 16S0E2 | ITS0E2 |
16S0F2 | ITS0F2 | |
MO1 | 16S0G2 | ITS0G2 |
16S0H2 | ITS0H2 | |
MO2 | 16S0A3 | ITS0A3 |
16S0B3 | ITS0B3 | |
ROCK1 | 16S0C3 | ITS0C3 |
16S0D3 | ITS0D3 | |
LVD1 | 16S0A11 | |
16S0B11 | ||
LVD2 | 16S0E11 | |
16S0F11 | ||
LVD3 | 16S0G12 | |
16S0H12 | ||
LVD4 | 16S0C4 | |
16S0D4 | ||
TT6 | 16S0G4 | |
16S0H4 | ||
TT7 | 16S0E3 | |
16S0F3 | ||
MO3 | 16S0E4 | |
16S0F4 | ||
AH1 | 16S0G3 | |
16S0H3 | ||
AH2 | 16S0A4 | |
16S0B4 |
qPCR and Sequencing
16S
Pool concentration = 2.87 nM
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