HyperMu™ <KAN-1> Insertion Kit
HyperMu™ <KAN-1> and <CHL-1> Insertion Kits are designed to simplify and speed up sequencing any cloned DNA over 2 kb. These kits use HyperMu™ MuA Transposase, a hyperactive mutant MuA transposase with an in vitro transposition efficiency at least 50 times higher than similar enzymes from other sources.
Like the EZ-Tn5™ Transposon Insertion Kits, the HyperMu Insertion Kits simplify and speed up complete sequencing of any cloned DNA that is too large to sequence with a single set of sequencing reactions.
The process for sequencing even the largest BAC clone without the time and expense of subcloning or primer walking is shown in Fig. 1. First, the HyperMu Transposon is randomly inserted into the target DNA using a simple, 2-hour in vitro reaction catalyzed by HyperMu Transposase. Then, following transformation of E. coli and plating on medium containing either kanamycin or chloramphenicol, up to a million independent insertion clones are obtained. The complete sequence of the target DNA is easily obtained by sequencing selected insertion clones bidirectionally using the two primers in the kit that are homologous to the ends of the inserted HyperMu Transposon.
|Figure 1 (click to enlarge). The process for generating DNA sequencing templates using a HyperMu™ Insertion Kit. Select inserts on kanamycin or chloramphenicol plates.|
- A single reaction generates enough insertion clones to sequence even the largest BAC clone.
- Use the sequencing primers provided in the kit to completely sequence your clone; no additional primer synthesis required.
- Insertion specificity is highly random, ensuring complete sequence coverage.