Template_amplicon_pooling.xlsx

	A	B	C	D	E	F	G	H	I	J	K	L	M	N	O	P	Q	R	S	T	U	V	W	X	Y	Z
1		Amplicon Pooling Template
2
3		1. Indicate the number of samples you have
4
5		Number of samples		59
6
7
8		2. Select the primers you used
9
10		16s	TRUE	16s	310	16s	1	10	10	16s	16s	310
11		18s	TRUE	18s	270	18s	2	11	11	18s	18s	270
12		FITS	TRUE	FITS	370	FITS	3	12	12	FITS	FITS	370
13		PITS	TRUE	PITS	520	PITS	4	13	13	PITS	PITS	520
14		CO1	TRUE	CO1	430	CO1	5	14	14	CO1	CO1	430
15		12S	FALSE	trnL						#REF!
16		16SMamm	FALSE	Other						#REF!
17
18
19		3. Indicate the PCR Product length and the max volume of sample you wish to use.
20		You can batch edit in Table 1, or modify on case by case basis in Table 2. You only need to modify/ verify cells highlighted in blue
21
22				Table 1. Batch editing
23				Primer	PCR Product Length	Max volume of sample you would like to add *		*(considering you eluted to a certain amount, subtracted 2 for quantification and should account for pippeting error- generally recommend 10)
24				16s	310	10
25				18s	270	10
26				FITS	370	10
27				PITS	520	10
28				CO1	430	10
29				12S	150	10
30
31
32
33		4. Add the concentrations calculated from the Qubit fluorescence to Table 2 below and verify the average amplicon length you observed in your gel.
34
35		5. How much DNA do you want to have in your indexing PCR (generally 5 or 10ng)
36
37		DNA for indexing PCR		50	ng
38
39		5. Look at results in the "Data output" tab.
40
41
42		Table 2. Detailed table
43				16s				18s				FITS				PITS				CO1				12S
44	Sample #	Enter your sample names here		PCR Product Length	Concentration (ng/uL)	Max volume of sample you would like to add (uL)		PCR Product Length	Concentration (ng/uL)	Max volume of sample you would like to add (uL)		PCR Product Length	Concentration (ng/uL)	Max volume of sample you would like to add (uL)		PCR Product Length	Concentration (ng/uL)	Max volume of sample you would like to add (uL)		PCR Product Length	Concentration (ng/uL)	Max volume of sample you would like to add (uL)		PCR Product Length	Concentration (ng/uL)	Max volume of sample you would like to add (uL)
45	(Example)	K0000_A1	16s	310	1.4638	10	18s	270	2.9638	10	FITS	370	1.4998	10	PITS	520	2.4738	10	CO1	430	4.7632	10	12S	150	1.48922	10
46	1		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
47	2		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
48	3		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
49	4		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
50	5		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
51	6		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
52	7		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
53	8		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
54	9		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
55	10		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
56	11		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
57	12		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
58	13		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
59	14		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
60	15		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
61	16		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
62	17		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
63	18		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
64	19		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
65	20		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
66	21		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
67	22		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
68	23		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
69	24		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
70	25		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
71	26		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
72	27		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
73	28		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
74	29		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
75	30		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
76	31		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
77	32		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
78	33		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
79	34		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
80	35		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
81	36		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
82	37		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
83	38		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
84	39		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
85	40		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
86	41		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
87	42		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
88	43		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
89	44		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
90	45		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
91	46		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
92	47		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
93	48		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
94	49		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
95	50		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
96	51		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
97	52		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
98	53		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
99	54		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10
100	55		16s	310		10	18s	270		10	FITS	370		10	PITS	520		10	CO1	430		10	12S	150		10