Contents > Transposomics™ > HyperMu™ <KAN-1> Insertion KitHyperMu™ <CHL-1> Insertion Kit
HyperMu™ <KAN-1> Insertion Kit
HyperMu™ <CHL-1> Insertion Kit
Supplier: EPICENTRE® Biotechnologies
HyperMu™ <CHL-1> Insertion Kit
| Catalogue number | Pack size |
| HMI039C | 10 standard reactions |
HyperMu™ <KAN-1> Insertion Kit
| Catalogue number | Pack size |
| HMI032K | 10 standard reactions |
HyperMu™ Transposase
| Catalogue number | Pack size |
| THM03210 | 10U |
Sterile Nuclease-Free Water
| Catalogue number | Pack size |
| W7350ML | 50ml |
TransforMax™ EC100™ Electrocompetent E. coli
| Catalogue number | Pack size |
| EC10005 | 5 x 100µl |
| EC10010 | 10 x 100µl |
TransforMax™ EC100™-T1R Electrocompetent E. coli
| Catalogue number | Pack size |
| EC0205T1 | 5 x 100µl |
 |
Get your Epicentre® Forum - hot off the press! Click here for direct link to Forum listings |
Last modified: 2008-07-15 14:57:15
Uses HyperMu™ Transposase, a hyperactive mutant MuA transposase with an in vitro transposition efficiency at least 50 times higher than similar enzymes from other sources, to facilitate complete sequencing of all your cDNA or genomic clones.
Like the EZ-Tn5™ Transposon Insertion Kits, the new 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. Epicentre®'s Mu-based transposition system uses a unique HyperMu™ MuA Transposase that retains the highly random insertion characteristics of MuA transposase but is at least 50 times more active in vitro than the MuA transposase from other suppliers. The higher activity of HyperMu Transposase results in much higher transposition efficiencies that are critical for obtaining a sufficient number of transposon insertions to completely sequence a clone, especially those with large inserts.
The process for sequencing even the largest BAC clone without the time and expense of subcloning or primer walking is shown in Figure 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. The process for complete sequencing of a target DNA using the HyperMu™ <KAN-1> Insertion Kit. |
Last modified: 2008-05-14 16:46:46
Applications
- In vitro insertion of an artificial Mu transposon into DNA cloned in vectors, such as plasmids, fosmids, cosmids, or BACs, as well as in vitro insertion into linear DNA.
- Preparation of HyperMu™ Transposomes for in vivo transposition of an artifical Mu transposon following electroporation into living cells.
Benefits
- Faster - completely sequence plasmid, cosmid, fosmid, and BAC clones 10-times faster than by subcloning or primer walking.
- Easier - a single reaction generates enough insertion clones to sequence even the largest BAC clone.
- More Economical - use only the sequencing primers provided in the kit to completely sequence your clone.
- Complete Sequence Coverage - insertion specificity is highly random to obtain the complete sequence of your DNA.
Last modified: 2008-05-14 16:46:46
Protocols for: HyperMu™ <KAN-1> & <CHL-1> Insertion Kit
HyperMu™ <KAN-1> Insertion Kit Protocol
(catalogue number HMI032K)
HyperMu™ <KAN-1> Transposon, MUKAN-1 FP-1 Forward primer & MUKAN-1 RP-1 Reverse Primer Protocol
(catalogue number MKAN320 / MKFP321 / MKRP322)
HyperMu™ <CHL-1> Insertion Kit Protocol
(catalogue number HMI039C)
HyperMu™ <CHL-1> Transposon, MUCHL-1 FP-1 Forward primer & MUCHL-1 RP-1 Reverse Primer Protocol
