NGS Multiplexing Simplified

Increased multiplexing to maximize sequencing

Multiplexing (e.g. the combining together of many samples that have been tagged with a unique sequence, called indexes) enables higher sequencing throughput and reduced sequencing cost by sequencing many more samples on the same sequencing run. Take full advantage of the increased sequencing capacity of Illumina sequencing instruments, such as the NovaSeq, with up to 192 pre-plated Unique Dual Index (UDI) adaptors!

 

Tecan's Universal Plus mRNA-Seq with NuQuant library preparation kit now comes standard with up to 192 pre-plated UDI adaptors.  

  • Maximize your sequencing capacity by multiplexing up to 192 samples
  • Increase your multiplexing flexibility
  • Unique pre-plated UDI adaptors for each sample included in the kit, no need to buy multiple sets of adaptors

What is index hopping?

Index hopping or index switching is a phenomenon that occurs during sequencing of a multiplexed library pool, where a sample index is mistakenly assigned to the wrong sample resulting in inaccurate assignment of reads.  This phenomenon is found to be associated with Illumina sequencing instrument at a low level and is elevated with patterned flow cells like those used with the Illumina NovaSeq instrument.  Additionally, the presence of a contaminating indexed primer or adaptor in the final library can increase the frequency of index hopping.  The Universal Plus mRNA-Seq kit workflow and UDI adaptors reduce the occurrence of index hopping and allow you to detect and eliminate inaccurately assigned reads.

 

Tecan's Universal Plus mRNA-Seq with NuQuant library preparation kit provides:

  • Unique UDI adaptors for each sample eliminating the need for multiple sample pools
  • Single-use pre-plated adaptors reduce the possibility of cross-contamination
  • Sample indexing during adaptor ligation reducing the possibility of adaptor contamination in the final library

Best practices to minimize index hopping

Index hopping is a consequence of multiplexing during sequencing on Illumina sequencing platforms. Best practices to minimize the impact of index hopping are listed below:

  1. Use Unique Dual Indexing to enable detection and elimination of inaccurately assigned sample reads - standard with Universal Plus mRNA-Seq with NuQuant kits!
  2. Careful cleanup of final libraries to minimize indexed primer carryover - Universal Plus mRNA-Seq with NuQuant does not use indexed primers for library amplification!
  3. Proper storage of unpooled libraries at -20 °C.
  4. Pool libraries immediately before sequencing.

Dual indexes vs. Unique Dual Indexes

Dual indexes provide the ability to increase the level of multiplexing while UDI adaptors allow you to detect index hopping. With 192 UDI adaptors, get the best of both worlds!

 

The Universal Plus mRNA-Seq with NuQuant library preparation kits come standard with up to 192 UDI adaptors.

ProductPart Number
Universal Plus mRNA-Seq with NuQuant0520-24, 0520-A01, 0520B-A01
Universal Plus mRNA-Seq with NuQuant, Human Globin AnyDeplete0521-24, 0521-A01, 0521B-A01
Universal Plus mRNA-Seq with NuQuant, Custom AnyDeplete0522-24, 0522-A01, 0522B-A01

 

 

 

 

 

 

For Research Use Only. Not for Use in Diagnostic Procedures.