1HILL

Prep

Extract samples using Qiagen Power Soil 96 kit

Quantify via absorbance

Normalize to 10 ng/ul on Nimbus

ul/rxn	Reagent	# of rxns	ul needed

ul/rxn	Reagent	# of rxns	ul needed
3	5X Kapa HiFi Buffer	850	2550
0.45	10M dNTPs	850	382
0.3	Kapa HiFi HotStart DNA Pol	850	255
5.25	HPLC H2O	850	4463
9	Total Volume	850	7650

Add 9 ul to each well of a hard shell, full skirt plate. Seal with bubble strips and store in refrigerator until needed.
Array primers into plates with Nimbus
Add 2 ul sample per well

Plate	16S Primer	ITS Primer

Pool concentration = 2.87 nM

1nM Dilution = 69.6 ul Pool + 130.4 ul RSB

Load = 90 ul 1 nM + 9 ul PhiX + 1 ul RSB