FAQ

What experiment should I do first to verify NTT-seq is working in my lab?

We recommend first running a bulk-cell experiment using validated H3K27me3 and H3K27ac antibodies. You should verify that you’re able to get good histone modification data for both marks with low signal mixing (low H3K27me3 signal in H3K27ac peaks and vice-versa).

How can I produce the nb-Tn5 protein?

You can get each nb-Tn5 plasmid from AddGene, and follow the Tn5 expression and purification protocol shown in Picelli et al. 2014.

How do I install a custom run recipe on my sequencer?

You will need to contact Illumina for support in adding a custom run recipe to your sequencer.

How do I load custom sequencing primers in a sequencing run?

You can either spike in custom primers (needed if you are sequencing standard libraries on the same run), or you can add custom primers to the run. Protocols for doing this vary depending on the sequencing instrument used, and the information can be found on the Illumina website.

How can I order custom sequencing primers?

See the reagents page for information.