Craig, Nancy (Johns Hopkins School of Medicine)
Gangadharan, Sunil (Johns Hopkins School of Medicine)
Dyda, Frederick (NIDDK)
Hickman, Allison (NIDDK)
This technology includes novel hyperactive Hermes Transposase mutants and their encoding genes. These transposases are easily purified in large quantity after expression in bacteria. The modified Hermes Transposases are soluble and stable and exist as smaller active complexes compared to the native enzyme. The consensus target DNA recognition sequence is the same as the native enzyme and shows minimal insertional sequence bias. These properties are especially useful in whole genome sequencing applications that involve sample DNA preparation requiring simultaneous fragmentation and attachment of custom sequences to the ends of the fragments. Methods and compositions using these transposases in fragmentation and 5' end-tagging are also disclosed.
Production and use in the generation of sequencing libraries.
- The transposases described in this invention have a similar mechanism of action as the wild type, can easily be expressed in the bacterium, E. coli and purified in large quantities.
- The modified Hermes Transposases have a higher transposition activity in vitro than the wild type transposase.
- The modified Hermes Transposase has less insertional sequence bias when used for in vitro fragmentation of genomic DNA and 51 end tagging followed by next generation sequencing.