Dear Chrystelle Delord

I haven't used this GT-seq method, but I got very good results using the Access Array platform from Fluidigm. This method allows up to 48 markers amplified in 48 samples (2,304 different amplicons) in one PCR. I used this method and it worked REALLY well for me, and it's a pretty cheap method, because you perform PCRs in a very small volume, so you use only 3 uL of Taq Polymerase for each plate (48 indsx48 markers).

This platform also allows you to multiplex up to primer pair in each primer well, so you could use up to 480 markers, but I haven't tried this multiplex implementation, so I can't tell you how good this method is.

I think it would be a good idea to check this strategy.

Hope this can be of any help,

Best,

Manolo

Thank you very much Manolo !

I had a look on this method, I don't know if this will be cheaper than performing target enrichment with classical PCR like in GT-seq, but this sure could be less tricky !

I am still open to other ideas :-) Thank you all !

Dear Chrystelle Delord

I have seen some multiple PCR methods which are similar to GT-seq.One method is Hi-Plex(Abridged adapter primers increase the target scope of Hi-Plex,2015).The author amplified 1000 amplicons in a single PCR tube and 94.12% of amplicons were represented at a coverage depth within 25-fold of the target-set median.I successfully amplified 56 amplicons(400bp) from 160 samples using the method and 76% amplcions fell within a 100-fold abundance range. Other methods  are described in the article (Multiplex PCR in Molecular Differential Diagnosis of Microbial Infections: Methods, Utility, and Platforms,2013), such as tem-PCR, tem-PCR, DPO and Nested Patch PCR. These methods can simultaneously amplify hundreds of amplicons.

Hope this can be of any help,

Best,

Ke

Thank you very much Ke ! This helps me a lot :)

Have a very nice day !

Cheers,

Chrys