Applications
- Completely sequence even the largest cDNA or genomic clones without
subcloning or primer walking.
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. |
