Dual index barcode demultiplex issue

Dual index barcode demultiplex issue

1

Hello,
Following are the list of dual indexed illumina libraries we have used for illumina run.But we were not able demultiplex the data by using the barcodes.Because 99% of the data going as undetermined.

Index1 —— Index2 ( case1 )

ATGAGC —- TGAACCTT

ATTCCT —- TGAACCTT

CAAAAG —- TGAACCTT

CAACTA —- TGAACCTT

CACCGG —- TGAACCTT

There after we tried demultiplex with reverse compliment of index 2 and No changes in Index1(list are below ).This worked well and able to determine 98% of the data.The question is why the demultiplex only worked with reverse compliment of index2 and not in the case1(without reverse compliment)

Index1 —— Index2 (reverse compliment)

ATGAGC —- AAGGTTCA

ATTCCT —- AAGGTTCA

CAAAAG —- AAGGTTCA

CAACTA —- AAGGTTCA

CACCGG —- AAGGTTCA


bcltofastq


Demultiplex


dual index barcod


hiseq4000

• 4.8k views

Source link