Indexing strategy and number of indexes: Including a short index sequence (8–10 bases)
in your adapters/primer mix
enables multiple samples to be sequenced together (i.e., multiplexed) on the same instrument flow cell or chip. Each sample index is specific to a given sample library and is used for de-multiplexing during data analysis to assign individual sequence
reads to the correct sample.
Select from barcode indexes of 8 or 10 bases, with sequences designed by IDT or Illumina (IDT 8, IDT 10, ILMN 8, ILMN 10). See the Resources tab below, for links to the IDT 8, IDT 10, and ILMN 8 index sequence files. The selected IDT
8, IDT 10, ILMN 8, or ILMN 10 index sequences will be provided once your Custom NGS Adapters order has been placed.
Alternatively, you can upload up to 2000 custom index sequences.
Choose to have the barcode indexes numbered consecutively or with numbering starting at predefined index positions.
Size (reaction number or nmol): Receive adapter amounts based on final mass (2, 8, or 24 nmol), or TruSeq/Nextera reaction number (16 or 96 reactions).
|Adapter or primers||Concentration (μM)||1 reaction (nmol)|
|TruSeq™–Compatible Full-length Adapters||15||0.075|
|TruSeq™–Compatible Stubby Adapter||15||0.075|
|Nextera™–Compatible Indexing Primers||10||0.050|
Delivery format: Select to have your adapters delivered in tubes, plates, or single-use plates. Adapters and primer mixes arrive on dry ice. TruSeq™–Compatible Full-length Adapters and the TruSeq™–Compatible Stubby
Adapter are suspended in Nuclease-free Duplex Buffer (30 mM HEPES, pH 7.5; 100 mM potassium acetate; IDT). TruSeq™–Compatible Indexing Primers and Nextera™–Compatible Indexing Primers are suspended in IDTE (1X TE Solution,
pH 8.0; IDT).
Methylated/unmethylated: Unmethylated adapters will suffice for most applications. Methylated adapters are used with epigenetics studies that require bisulfate conversion.
TruGrade processing: Sensitive NGS applications can detect even low levels of barcode crosstalk. Barcode crosstalk can cause sample misassignment. There are multiple sources of barcode crosstalk, including manufacturing cross-contamination
of adapter oligos. By including TruGrade processing, you can reduce levels of cross-contamination to as low as 0.01%.
Find more details on the IDT TruGrade webpage and in the DECODED Newsletter article, Tips for minimizing sample misassignment during multiplex NGS.