Skip to main content

Table 3 Sequences of primers used for MiSeq sequencing

From: 16S rRNA gene sequencing of mock microbial populations- impact of DNA extraction method, primer choice and sequencing platform

Sample

Primer sequence

Barcode

Ref

V4-V5 primer

  

[23]

 Forward primer

AATGATACGGCGACCACCGAGATCTACACTATGGTAATTGGGTGCCAGCMGCCGCGGTAA

  

 Read 1 primer

TATGGTAATTGGGTGCCAGCMGCCGCGGTAA

  

 Read 2 primer

AGTCAGTCAGTTCCGTCAATTYYTTTRAGTTT

  

 Index primer

AAACTYAAARRAATTGACGGAACTGACTGACT

  

Reverse barcoded primers

 PBS Qiagen

CAAGCAGAAGACGGCATACGAGATTAACGTGTGTGCAGTCAGTCAGTTCCGTCAATTYYTTTRAGTTT

TAACGTGTGTGC

 

 PBS RBB

CAAGCAGAAGACGGCATACGAGATCATTATGGCGTGAGTCAGTCAGTTCCGTCAATTYYTTTRAGTTT

CATTATGGCGTG

 

 Qiagen Glycerol

CAAGCAGAAGACGGCATACGAGATCCAATACGCCTGAGTCAGTCAGTTCCGTCAATTYYTTTRAGTTT

CCAATACGCCTG

 

 RBB Glycerol

CAAGCAGAAGACGGCATACGAGATGATCTGCGATCCAGTCAGTCAGTTCCGTCAATTYYTTTRAGTTT

GATCTGCGATCC

 

 Mock DNA

CAAGCAGAAGACGGCATACGAGATCAGCTCATCAGCAGTCAGTCAGTTCCGTCAATTYYTTTRAGTTT

CAGCTCATCAGC

 

V1-V2 set 1

  

[4]

 Forward primer

AATGATACGGCGACCACCGAGATCTACACTATGGTAATTTCAGAGTTTGATCCTGGCTCAG

  

 Read 1 primer

TATGGTAATTTCAGAGTTTGATCCTGGCTCAG

  

 Read 2 primer

AGTCAGTCAGCATGCTGCCTCCCGTAGGAGT

  

 Index primer

ACTCCTACGGGAGGCAGCATGCTGACTGACT

  

Reverse barcoded primers

 PBS Qiagen

CAAGCAGAAGACGGCATACGAGATTCTTGGAGGTCAAGTCAGTCAGCATGCTGCCTCCCGTAGGAGT

TCTTGGAGGTCA

 

 PBS RBB

CAAGCAGAAGACGGCATACGAGATTCACCTCCTTGTAGTCAGTCAGCATGCTGCCTCCCGTAGGAGT

TCACCTCCTTGT

 

 Qiagen Glycerol

CAAGCAGAAGACGGCATACGAGATGCACACCTGATAAGTCAGTCAGCATGCTGCCTCCCGTAGGAGT

GCACACCTGATA

 

 RBB Glycerol

CAAGCAGAAGACGGCATACGAGATGCGACAATTACAAGTCAGTCAGCATGCTGCCTCCCGTAGGAGT

GCGACAATTACA

 

 Mock DNA

CAAGCAGAAGACGGCATACGAGATTCATGCTCCATTAGTCAGTCAGCATGCTGCCTCCCGTAGGAGT

TCATGCTCCATT

 

V1-V2 degenerate

 

[17]

 Forward primer

AATGATACGGCGACCACCGAGATCTACACTATGGTAATTTCAGMGTTYGATYMTGGCTCAG

  

 Read 1 primer

TATGGTAATTTCAGMGTTYGATYMTGGCTCAG

  

 Read 2 primer

AGTCAGTCAGCATGCTGCCTCCCGTAGGAGT

  

 Index primer

ACTCCTACGGGAGGCAGCATGCTGACTGACT

  

Reverse barcoded primers

 PBS Qiagen

CAAGCAGAAGACGGCATACGAGATTCTTGGAGGTCAAGTCAGTCAGCATGCTGCCTCCCGTAGGAGT

TCTTGGAGGTCA

 

 PBS RBB

CAAGCAGAAGACGGCATACGAGATTCACCTCCTTGTAGTCAGTCAGCATGCTGCCTCCCGTAGGAGT

TCACCTCCTTGT

 

 Qiagen Glycerol

CAAGCAGAAGACGGCATACGAGATGCACACCTGATAAGTCAGTCAGCATGCTGCCTCCCGTAGGAGT

GCACACCTGATA

 

 RBB Glycerol

CAAGCAGAAGACGGCATACGAGATGCGACAATTACAAGTCAGTCAGCATGCTGCCTCCCGTAGGAGT

GCGACAATTACA

 

 Mock DNA

CAAGCAGAAGACGGCATACGAGATTCATGCTCCATTAGTCAGTCAGCATGCTGCCTCCCGTAGGAGT

TCATGCTCCATT

Â