| Info |
|---|
Samples arrived 05-05-2022. Samples checked in against list 7-13-2022. Plates 1-3 have been extracted, normalized, PCRed, quantified and normalized by 7-15-22. 16S for 2TRNL1 was re-PCRed with G4 and H4 because there was low amplification with A5 and B5. |
Extractions
- Transfer fecal material to labeled bead tubes. Weigh first few samples to approximate amount of material ~200mg
- Freeze dry samples
- Extract using MN Nucleospin 96 Stool kits following spin speeds from MN Nucleospin 96 Soil kits
- Transfer DNAs to plate
- Transfer 30 ul as working aliquot
- Add 4 samples of 5 ng/ul Mock Community Only and 4 samples of half mock community half DNA from a single plant sample DNA to column 9(first empty column) of 2TRNL8
- Add ISD and coligos
- Normalize
...
ul/rxn | Reagent | # of rxns | ul needed |
|---|---|---|---|
3 | 5X Kapa HiFi Buffer | 19002690 | 58,700070 |
0.45 | 10M dNTPs | 19002690 | 8551211 |
0.3 | Kapa HiFi HotStart DNA Pol | 19002690 | 570807 |
7.25 | HPLC H2O | 19002690 | 1319,775502 |
11 | Total Volume | 19002690 | 2029,900590 |
Add 11 ul to each well of a hard shell, full skirt plate. Add 2 uL of primers and 2uL of template to each well.
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