Plate | 16S Primer | ITS Primer | |
|---|---|---|---|
6LVD5 | 16S0A4 | ITS0A4 | |
16S0B4 | ITS0B4 | ||
6LVD66RD4 | 16S0C4 | ITS0C4 | |
16S0D4 | ITS0D4 | ||
6LVD76RD5 | 16S0E4 | ITS0E4 | |
16S0F4 | ITS0F4 | ||
6RD6 | 16S0C5 | ITS0C5 | |
16S0D5 | ITS0D5 | ||
6RD7 | 16S0E5 | ITS0E5 | |
16S0F5 | ITS0F5 |
MasterMix (make for 16 plates in 50mL conical tube)
ul/rxn | Reagent | # of rxns | ul needed |
|---|---|---|---|
3 | 5X Kapa HiFi Buffer | 1400 | 4200 |
0.45 | 10M dNTPs | 1400 | 630 |
0.3 | Kapa HiFi HotStart DNA Pol | 1400 | 420 |
7.25 | HPLC H2O | 1400 | 10,150 |
11 | Total Volume | 1400 | 15400 |
...
Temp C | Cycles | Time |
|---|---|---|
95* | 1X | 3:00* |
98 | 36X | 0:30 |
62 | 36X | 0:30 |
72 | 36X | 0:30 |
72 | 1X | 5:00 |
4 | 1X | 0:00 |
MagBead Cleanup:
Some plates were cleaned up using the Nimbus platform protocol “AxyPrep MagBead PCR1 No MM”
...