Effects of polymerase, template dilution and cycle number on PCR based 16 S rRNA diversity analysis using the deep sequencing method

Table 1 Sample list

ID	Barcode	PCR conditions			Read number		Chao1		Ace
		T*	C^&	E^$	(total)	(unique)	(unique)	0.03	(unique)	0.03
A1	TGGAGTAG	1	30	Ex	83,194	17,841	58,148	13,020	108,316	18,590
A2	TGTGACTG	1	30	Ex	158,519	30,361	55,899	34,096	107,984	22,871
B1	CAGACAGA	20	30	Ex	52,793	12,874	39,159	7,455	69,614	9,274
B2	CAGTGAGA	20	30	Ex	78,392	16,846	50,838	8,986	88,268	10,782
C1	CATCTCGT	200	30	Ex	25,705	6,013	16,586	2,700	24,554	2,669
C2	GGTAGGAT	200	30	Ex	25,514	5,968	16,828	2,731	25,294	2,649
D1	GTGTAGAG	20	25	Ex	10,833	3,992	13,749	4,457	26,155	6,406
D2	GTTGGTAC	20	25	Ex	25,181	7,578	22,921	6,698	42,784	9,517
E1	GTCAGAGA	20	30	Pfu	34,600	6,750	17,853	6,332	30,589	9,255
E2	GTCTTCTG	20	30	Pfu	35,152	6,818	18,281	6,416	30,434	8,792
	Total				529,883	67,826	229,287	34,883	120,750	50,579

*: Dilution folds of the DNA template; &: PCR cycle number; $: Polymerase used (Ex, Ex Taq from Takara; Pfu, PfuUltra II Hotstart 2× Master Mix from Stratagene).

ISSN: 1471-2180