16S rRNA gene primer choice impacts off-target amplification in human gastrointestinal tract biopsies and microbiome profiling

Table 1 Primers used for 16S rRNA gene sequencing analysis.

16S region	Name	c (µmol L⁻¹)	Sequence (5′–3′)
V4	515F³⁶	0.2	5′AATGATACGGCGACCACCGAGATCTACACGCTGCGTAAGATATGGTAATTGTGTGYCAGCMGCCGCGGTAA 3′
	806R³⁶	0.2	5′CAAGCAGAAGACGGCATACGAGATTCGCCTTAAGTCAGCCAGCCGGACTACNVGGGTWTCTAAT 3′
	Read1		TATGGTAATTGTGTGCCAGCMGCCGCGGTAA
	Read2		AGTCAGTCAGCCGGACTACHVGGGTWTCTAAT
	Index1		ATTAGAWACCCBDGTAGTCCGGCTGACTGACT
	Index2		TTACCGCGGCKGCTGGCACACAATTACCATA
V1–V2	68F¹⁶	0.2	5′AATGATACGGCGACCACCGAGATCTACACGCTCGTCTAATAGTCAGCCAGCCGTNANACATGCAAGTCGRRSG 3′
	68F_M	0.1	5′AATGATACGGCGACCACCGAGATCTACACGCTCGTCTAATAGTCAGCCAGCCGTAACACATGCAAGTCRACTYGA 3′
	338R¹⁷	0.3	5′CAAGCAGAAGACGGCATACGAGATTCATGAGCTATGGTAATTAAGCTGCCTCCCGTAGGAGT 3′
	Read1		AGTCAGCCAGCCGTNANACATGCAAGTC
	Read2		TATGGTAATTAAGCTGCCTCCCGTAGGAGT
	Index1		ACTCCTACGGGAGGCAGCTTAATTACCATA
	Index2		GACTTGCATGTNTNACGGCTGGCTGACT