Golay Barcodes vs. Nextera Sequencing Adapter Barcodes for Mapping File

[slh277]

Hi, thank you all so much for your continued support and help!!!

I am having trouble determining how to make my metadata mapping file, for paired-end reads. The company used dual-indexing Nextera Adapters and didn’t specify the Golay barcodes named as such, just the i5 and i7 index sequences (the data came to us demultiplexed and trimmed already). I wasn’t sure how to incorporate Nextera (or what barcode to use) into my mapping file (draft below):

#SampleID BarcodeSequence LinkerPrimerSequence ReverseLinkerPrimerSequence
5.105-G ? GTGTGYCAGCMGCCGCGGTAA CCGGACTACNVGGGTWTCTAAT
5.111-B ? GTGTGYCAGCMGCCGCGGTAA CCGGACTACNVGGGTWTCTAAT

The Nextera info would be -

#Sample ID Index 1 [i7] Index 1 sequence Index 2 [i5] Index 2 sequence
5.105-G N723 TAGCGCTC S515 TTCTAGCT
5.111-B N702 CGTACTAG S513 TCGACTAG

Any help would be much appreciated!

Thank you,
Sam

[thermokarst]

Hi @slh277 — if the sequences are already demultiplexed, you don’t need to worry about including your barcodes in your metadata file - that is only necessary if you are going to use QIIME 2 to demultiplex your reads. Hope that helps!