difference between target sequencing technologies

Question

[pingu77]

Hello!

What is the difference between hybrid capture sequencing and whole exome sequencing? It seems that both are target sequencing technology and I cannot understand the difference.

Thank you in advance for your help

Answer 1

[GenoMax]

Target capture is a generic method for enrichment of specific regions (which can include coding genes). There are other applications that use this as well : https://www.idtdna.com/pages/technology/next-generation-sequencing/dna-sequencing/targeted-sequencing/hybridization-capture

Whole exome sequencing on other hand is just looking at protein-coding regions with target capture being the method (in solution or on arrays) used for enrichment: https://en.wikipedia.org/wiki/Exome_sequencing

[pingu77]

Thank you GenoMax. If I understand correctly, it is possible to capture non-coding regions with hybridization technology, right? Are there any limitations with hybridization capture technology? (Other than repeated regions, which they say can be overcome with probe design) Last question (sorry I don't have wet lab experience at all): is hybrid capture sequencing capable of sequencing a full gene? The link you sent states "Regions of interest within the library are then captured using biotinylated oligonucleotide probes", but how extensive could the regions be?

Thank you!

[GenoMax]

Are there any limitations with hybridization capture technology?

Yes you should be able to capture any region subject to proper probe design. There is a cost for synthesizing these probes.

is hybrid capture sequencing capable of sequencing a full gene?

In theory yes. You can have probe designs that tile the entire gene. see: https://www.takarabio.com/learning-centers/next-generation-sequencing/technical-notes/rna-seq/full-length-mrna-seq-for-target-capture

[pingu77]

Thanks a lot for your help!