Question: Demultiplexing fastq without barcode; with QIIME
0
gravatar for nidhiv
2.7 years ago by
nidhiv0
nidhiv0 wrote:

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 • 1.2k views
ADD COMMENTlink modified 2.7 years ago by Brian Bushnell17k • written 2.7 years ago by nidhiv0

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)

ADD REPLYlink modified 2.7 years ago • written 2.7 years ago by Chris Fields2.1k
2
gravatar for Brian Bushnell
2.7 years ago by
Walnut Creek, USA
Brian Bushnell17k wrote:

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.

ADD COMMENTlink written 2.7 years ago by Brian Bushnell17k
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