TLA for Cells & HMW DNA

TLA for Cells & HMW DNA

The Targeted Locus Amplification (TLA) technology uniquely enables targeted complete sequencing of genes of interest. In the protocol such as published in Nature Biotechnology, TLA consists of the following steps:

  • DNA crosslinking
  • DNA fragmentation
  • Religation and decrosslinking yields circular TLA
  • Stochastic reshuffling in millions of cells
  • Inverse PCR

 

Result of TLA

As a result, the complete locus is amplified and can be sequenced using Next Generation Sequencing technologies.

 

The TLA protocol results in the enrichment and sequencing of large regions surrounding the short locus specific sequences used for targeted enrichment. Since the TLA technology exclusively relies on physical proximity as basis of selection, it uniquely enables the targeted complete sequencing of regions of interest and detection of both all single nucleotide variants (SNVs) and structural variants (SVs).

TLA can be performed on cells and, with some modifications in the first steps, HMW DNA.

 

The TLA protocol can be executed manually, PerkinElmer has standardized and tested protocols automating the TLA protocol on the Sciclone® G3 NGSx workstation and Zephyr® G3 NGS workstation.

In addition, as a result of the reshuffling of DNA sequences originating from individual alleles that the protocol results in, TLA in combination with both paired-end and long read sequencing enables the haplotyping* of regions of interest. TLA can also be combined with linear PCR or capture (also see TLA technology for FFPE).

* Cergentis does not currently offer this as a routine service.

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