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Version	Old Version 3	New Version 4
Changes made by	Gregg Randolph	Gregg Randolph
Saved on	Aug 03, 2022	Aug 04, 2022

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Make 1:1000 dilutions of column 1,6,10 4 or 2 and 4 (2AMF2) from the PCR plates by adding 1 ul to 999 ul TE in a deep well plate:

	1	2	3	4	5	6	7	8	9	10	11	12
A	2AMF1_AMF	2AMF1_ITS	2AMF2_AMF	2AMF2_ITS						NTC	NTC	NTC
B	2AMF1_AMF	2AMF1_ITS	2AMF2_AMF	2AMF2_ITS						0.0002 pM Std	0.0002 pM Std	0.0002 pM Std
C	2AMF1_AMF	2AMF1_ITS	2AMF2_AMF	2AMF2_ITS						0.002 pM Std	0.002 pM Std	0.002 pM Std
D	2AMF1_AMF	2AMF1_ITS	2AMF2_AMF	2AMF2_ITS						0.02 pM Std	0.02 pM Std	0.02 pM Std
E	2AMF1_AMF	2AMF1_ITS	2AMF2_AMF	2AMF2_ITS						0.2 pM Std	0.2 pM Std	0.2 pM Std
F	2AMF1_AMF	2AMF1_ITS	2AMF2_AMF	2AMF2_ITS						2 pM Std	2 pM Std	2 pM Std
G	2AMF1_AMF	2AMF1_ITS	2AMF2_AMF	2AMF2_ITS						20 pM Std	20 pM Std	20 pM Std
H	2AMF1_AMF	2AMF1_ITS	2AMF2_AMF	2AMF2_ITS

Add 16 ul of Illumina Library Quantification MasterMix to each well:

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Add 4 ul of template, pool, or standards to each well:

	1	2	3	4	5	6	7	8	9	10	11	12
A	BS2AMF1_PLT1AMF	2AMF1_Col1ITS	BS2AMF2_PLT1_Col6AMFBS	2TRNL_PLT1_Col10	BS_PLT2_Col1	BS_PLT2_Col6	BS_PLT2_Col10	BS_AMF_Pool		NTC16S	40 16S	40 16S	40 16S	40 ITS	40 ITS	40 ITS	NTC	NTC
B	BS2AMF1_PLT1_Col1AMFBS	2AMF1_PLT1_Col6ITSBS	2AMF2_PLT1_Col10AMFBS	2TRNL_PLT2_Col1	BS_PLT2_Col6	BS_PLT2_Col10	BS_AMF_Pool			0.0002 pM Std16S	4 16S	4 16S	4 16S	4 ITS	4 ITS	4 ITS	0.0002 pM Std	0.0002 pM Std
C	BS2AMF1_PLT1_Col1AMFBS	2AMF1_PLT1_Col6ITSBS	2AMF2_PLT1_Col10AMFBS	2TRNL_PLT2_Col1	BS_PLT2_Col6	BS_PLT2_Col10	BS_AMF_Pool			0.002 pM Std16S	0.4 16S	0.4 16S	0.4 16S	0.4 ITS	0.4 ITS	0.4 ITS	0.002 pM Std	0.002 pM Std
D	BS2AMF1_PLT1_Col1AMFBS	2AMF1_PLT1_Col6ITSBS	2AMF2_PLT1_Col10AMFBS	2TRNL_PLT2_Col1	BS_PLT2_Col6	BS_PLT2_Col10				0.02 pM StdT	0.04 16S	0.04 16S	0.04 16S	0.04 ITS	0.04 ITS	0.04 ITS	0.02 pM Std	0.02 pM Std
E	BS2AMF1_PLT1_Col1AMFBS	2AMF1_PLT1_Col6ITSBS	2AMF2_PLT1_Col10ITSBS	2TRNL_PLT2_Col1	BS_PLT2_Col6	BS_PLT2_Col10				0.2 pM StdT	0.004 16S	0.004 16S	0.004 16S	0.004 ITS	0.004 ITS	0.004 ITS	0.2 pM Std	0.2 pM Std
F	BS2AMF1_PLT1_Col1AMFBS	2AMF1_PLT1_Col6ITSBS	2AMF2_PLT1_Col10ITSBS	2TRNL_PLT2_Col1	BS_PLT2_Col6	BS_PLT2_Col10				2 pM StdT	0.0004 16S	0.0004 16S	0.0004 16S	0.0004 ITS	0.0004 ITS	0.0004 ITS	2 pM Std	2 pM Std
G	BS2AMF1_PLT1_Col1AMFBS	2AMF1_PLT1_Col6ITSBS	2AMF2_PLT1_Col10	BS_PLT2_Col1	BS_PLT2_Col6	BS_PLT2_Col10	ITS									20 pM Std	20 pM Std	20 pM Std
H	BS2AMF1_PLT1_Col1AMFBS	2AMF1_PLT1_Col6ITS	BS_PLT1_Col10	BS_PLT2_Col1	BS_PLT2_Col6	BS_PLT2_Col10	2AMF2_ITS

Results:

View file

name	BS_AMF.csv

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