TABLE 3.

Reaction setup and thermocycler settings used for experimental PCR

Parameter	Value of the parameter for:
	Rxn_1a	Rxn_2	Rxn_3	Rxn_4
Total vol (μl)	50	50	50	25
Primers	TR3-TR4	341f-785r	341f-785r	Barcodes
Master mix (μl)	20	20	20	10
10 μM primer (μl)	2	2	2	0.94
Template DNA (μl)	2	2	2	1.25
Salinibacter ruber DNA (μl)			0.59
Nuclease-free H2O (μl)	26	26	25.41	12.81
Initial denaturation
Temp (°C)	95	94	94	94
Duration (min)	2	3	3	1.5
Denaturation
Temp (°C)	95	94	94	94
Duration (min)	1	0.75	0.75	0.5
Annealing
Temp (°C)	55	60	60	60
Duration (min)	1.5	1	1	0.5
Elongation
Temp (°C)	72	72	72	72
Duration (min)	1.5	1.5	1.5	1.5
Total no. of cycles	35	35	35	10
Final elongation
Temp (°C)	72	72	72	72
Duration (min)	10	10	10	5

^aRxn, reaction.