Demultiplexing fastq without barcode; with QIIME

I have received a single forward and reverse fastq file for 21 separate sample data generated by Illumina Miseq. The sequence appears to have the barcodes removed.

@M04771:133:000000000-B59PL:1:1101:16445:1361 1:N:0:0
TCCTTTTTTTTCTCTCTTTTTTCTTCTTTCTTTTTTTTCCCTTTTTCTTCTTTTTTTTTTTCCTTCTTTTTTTCCCTCTTTTTTCTTCCCTTTTTTTTTTCTTTTTCTCCTTTTTTTCTTTTTTTTTTTTTTTTTTTTTTTCCTTTTTTTTTTCTCTTTTTCCTTCTTCTCTTTTTTTTTTCCTCCCTTTTTCTTCTTCTTTTTTCTTTTTTTTTTCTTCCTCTTCTTTTCTTCCTCTTTTTTCTCTCCCT
+
A>>>>31DAA>01AFFGFG110BGHG2FA22BDFG/AE011F1B110D1DFF1DF>////>01>>111>1BEE0B1B0011>1<<B1210<1111///<<011<110<00=00<0-::;000::?-9-9---99--99;-@//99///--9@@B9///9F///9://;///;/9//-9--;/////--/99:/;///9/9//9/-/999//--9--/99////9///////99/////99;99/;/////-

Any suggestions as to how I go about separating it to each sample? Preferably with QIIME

Thanks!

sequencing

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updated 9.1 years ago by Brian Bushnell

• written 9.1 years ago by nidhiv •

What Brian Bushnell says. You are missing the separate barcode FASTQ file which is required for demultiplexing (the -b option for split_libraries_fastq.py)

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written 9.1 years ago by Chris Fields

1 answer

You need to obtain the original data; it is impossible with what you have (at least, with the information given). In special cases (such as when each is a distinct, unrelated organism, for which you have a complete reference) there are techniques you can use to split the file, but you should still get the original data.

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written 9.1 years ago by Brian Bushnell