Description

Multiplexing Unique Dual Index Primers (Illumina Platform)

The Multiplexing Unique Dual Index Primers contains primer mix for multiplexing samples for Next Generation Sequencing (NGS) on the illumina platform. Multiplexing of NGS samples will reduce sequencing costs by pooling multiple NGS libraries into a single flow cell lane.

We have developed a Four-Base Difference Index System. The system allows us to make indexes that have at least 4 bases different from each other in the 8 bases index length. Our unique dual indexing primers remove sequencing errors such as index hopping, index cross-contamination, mis-assignment of reads, amplification errors, and de-multiplexing errors. The primer set includes 96 pre-mixed unique pairs of i5 and i7 index primers in a 96-well plate (index information can be downloaded Here).

Features

• • • Improves sample-identification specificity with 4-Base Difference Index System
      • Each index has at least 4 bases different from other indexes
      • 4-base difference greatly increases specificity compared to the other vendors’ 3-base difference.
    • Increases multiplexing capacity
      • 96 pre-mixed unique pairs of i5 and i7 index primers
    • Minimizes sequencing errors such as:
      • Index hopping
      • Index cross-contamination
      • Mis-assignment of reads
      • Amplification errors
      • De-multiplexing errors

Even distribution of 96 samples using dual index primers. 96 libraries were made using the BioDynami NGS DNA Library Prep Kit (Cat. # 30009) and the BioDynami Multiplexing Unique Dual Index Primers (Cat. # 30075). Libraries were pooled at equal concentration and sequenced on the illumina HiSeq 4000. The numbers of reads from 96 libraries were analyzed.

Identification of index hopping by unique dual index. 96 libraries were made using the BioDynami NGS DNA Library Prep Kit (Cat. # 30009) and the BioDynami Multiplexing Unique Dual Index Primers (Cat. # 30075). Libraries were pooled at equal concentration and sequenced on the illumina HiSeq 4000. The numbers of reads from all libraries were demultiplexed and analyzed.

• • • Demultiplexed: correct i5 and i7 index combination
    • Identified index hopping: correct barcodes, but NOT correct i5 and i7 index combination
    • Other: not fitting into the above categories

Sequence of the final library with index locations:

Related Products

• NGS DNA Library Prep Kit
• NGS DNA Library Prep Customization Kit
• PCR-free NGS DNA Library Prep Kit
• NGS Low Input DNA Library Prep Kit
• ChIP-Seq Library Prep Kit
• NGS Cell-free DNA Library Prep Kit
• NGS FFPE DNA Library Prep Kit
• Bisulfite Sequencing Library Prep Kit
• Methylation Specific Bisulfite-Seq Library Prep Kit
• NGS Single Stranded DNA Library Prep Kit
• NGS Ancient DNA Library Prep Kit
• NGS DNA Fragmentation & Library Prep Kit
• RNA Seq Library Prep Kit
• Multiplexing Index Primers (Illumina Platform)
• Multiplexing Unique Dual Index Primers (Illumina Platform)