What is the Tapestri barcode structure?

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Tapestri libraries support two barcode structures based on chemistry type – V1 and V2. V1 is the older chemistry type, and V2 is the latest version. Runs for the two differ in manufacturing, biochemistry, panel content, and barcode structure. V1 chemistry is not manufactured anymore.

A barcode is a unique DNA sequence ligated to fragments within a sequencing library for downstream in-silico sorting and identification. The barcodes are attached to the 5' end of the R1 (forward) reads. After generating the DNA libraries, these barcodes need to be removed from the sequence and used as cell identifiers.

For more information, refer to the Tapestri Pipeline User Guide.

V1 Chemistry

V1.png

Barcoding Bead (= Forward Primer): PhotoC-Element---Custom Adapter (33 bp)---BC1 (8 bp)---Constant (22 bp)---BC2 (8 bp)---Constant2 (8 bp)---Fwd primer (17 – 33 bp)

PhotoC-E is a linker that is bound between the Custom Adapter and the beads. This allows us to cleavage the oligo from the beads when exposed to long-wave, low-wattage UV light. (Integrated DNA Technologies). 

Custom Adapter (33) is a custom sequence, serving as the P5 primer annealing site when sequencing the library and requires a custom sequencing primer.

Reverse Primer: Rev primer (17 – 33 bp)---Read2 (34 bp)

V2 Chemistry

V2.png

Barcoding Bead: PhotoC-Element---Read 1 (36 bp)---BC1 (9 bp)---Off-Constant (14 – 17 bp)---BC2 (9 bp)---Constant2 (15 bp)

PhotoC-E is a linker that is bound between the Read 1 and the beads. This allows us to cleavage the oligo from the beads when exposed to long-wave, low-wattage UV light. (Integrated DNA Technologies). 

Read 1 is the read primer where Illumina’s sequencing primer binds to start the sequencing process.

With Off-Constant offsets of 0 to 3 as follows:

  • 0 Bases = no base
  • 1 Base = C
  • 2 Bases = TC
  • 3 Bases = GTC

Forward Primer: Constant2 (15 bp)---Fwd primer (17 – 33 bp)

Reverse Primer: Rev primer (17 – 33 bp)---Read2 (34 bp)

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