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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
  • DNA religation
  • Reversal of crosslinking
  • DNA circularisation
  • Inverse (multiplex) PCR

 

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. For more information please contact sales.

 

 

Compare technologies

See how TLA compares to conventional approaches such as WGS and FISH.

Terminology & methods

Read our manual on the terminology and methods used in TLA analyses.

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