1 reply to this topic

[JTS]

Gretings from New Zealand

I was wondering If any one has had any experience with in vitro transposition kits for sequencing. The two I am looking at are the epicenter EZ::Tn (Tn5 based), and the NEB GPS-1 system (Tn7 based). Are there any others worth considering, I've looked at the applied biosystems primer island (mariner-based) but thi sis pretty expensive. I'd be particularly interested if any one has had problems with non random insertions in repetitive DNA targets.

Many Thanks

John

[Yingjie]

hey, John

I am now using EZ::TN insertion kit for randonly mutagenesis of H.influenzae. I got quite a lot random insertions confirmed by southern blot. However, I couldn't sequence my genomic DNA template using the Tn5 outward primer.

Mariner is a better choice for random insertion purpose because it only recognizes TA dimer sequence. Tn5 recognizes 9bp sequence, while Tn7 has a hot recognization spot.

Good luck for your experiments.

Yingjie