Utilize the tremendous sequencing capacity of SOLiD® System with library multiplexing.

Reach Your DNA Sequencing Goals with Maximum Efficiency

SOLiD® DNA Barcoding Kits are currently available for SOLiD® fragment libraries.

SOLiD® Fragment Library Barcoding kits allow SOLiD® System users to multiplex up to 96 library samples on a single slide with the use of molecular barcodes. Multiplexing enables the pooling of samples prior to emulsion PCR (ePCR) on the SOLiD® System, which enables significant decreases in the cost and handling requirements of ePCR, enrichment, and deposition by allowing more samples per run. Such capabilities substantially benefit applications such as whole genome sequencing of small genomes or targeted resequencing of enriched samples.

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Figure 1. Barcoded fragment library design. Click to enlarge.

Molecular barcodes are 10-base sequences  embedded within the SOLiD® system sequencing adaptors (Figure 1), which are used to match sequence reads to original samples when read by the SOLiD® system. These specific DNA sequences are ligated directly to respective fragment libraries. The SOLiD® Fragment Library Barcoding Modules provide the necessary reagents to barcode fragment libraries from 8 µg input genomic DNA when used with SOLiD® Fragment Library Construction Kits.

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Figure 2. Click to enlarge.

Since SOLiD® Fragment Library Barcoding Kits follow the SOLiD® Fragment Library Construction protocol, it is easy to integrate barcodes in your fragment library construction workflow (figure 2).  With the availability of 6 modules, each containing 16 unique barcodes, it is possible to follow this workflow for up to 96 samples.

Figure 2. The SOLiD® Fragment Library Barcoding Workflow. Sheared DNA is end-repaired and purified for adaptor ligation.  At adaptor ligation, the fragmented, double-stranded DNA is ligated with a tuncated Multiplex P1 Adaptor and a Multiplex P2 Adaptor with a barcode.  The multiplex P2 Adaptor consists of three segments: and internal adaptor sequence, a barcode decamer sequence, and a standard P2 adaptor sequence.  Following adaptor ligation, correctly sized product is selected from the ligated DNA, which then undergoes nick-translation and amplification. The purified library is then quantitated via qPCR (Cat No. 4449639 SOLiD® Library TaqMan® Quantitation Kit) and pooled with other barcoded fragment libraries.

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Figure 3.

Figure 3. 96-multiplex Barcode Mapping Statistics to F3 Target Insert in E. Coli. End-repaired E. coli DH10B gDNA was split into 96 portions, each ligated to a different barcoded adaptor using SOLiD® DNA Barcoding Kits. The resulting libraries were amplified, then pooled for emulsion PCR and SOLiD® Sequencing. Target insert reads were binned by barcode, and mapped to the E. coli reference genome. Barcoding with the use of SOLiD® Fragment Library Barcoding Kits exhibits good mapping rates for the barcode and the DNA insert reads.