New England Biolabs, Inc.

Margaret Heider

Ming-Qun Xu

James C Samuelson

Minyong Chen

Aihua Zhang

Pingfang Liu

Lynne Apone

Brittany S Sexton

Louise J S Williams

Eileen T Dimalanta

Ming-Oun Xu

Provided herein is a polymerase-free enzyme mix (FRAG) for fragmenting double-stranded DNA. In some embodiments the enzyme mix may comprise a double-stranded DNA nickase and at least one of a DNA ligase capable of sealing a nick within a DNA, and a single-strand specific DNA nuclease. Methods for fragmenting double-stranded DNA are also provided.

Fragmentation of DNA

Provided herein is a thermolabile proteinase and methods of using the same. In some embodiments, the thermolabile proteinase may comprise an amino acid sequence that is at least 90% identical to any of SEQ ID NOs:1-11 and at least one amino acid substitution in helix 3. The thermolabile proteinase is active at a temperature in the range of 4&#xb0; C.-40&#xb0; C. and is inactivated by raising the temperature to above 50&#xb0; C., where the proteinase is substantially inactive at 65&#xb0; C.

Proteinases with improved properties

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Ipswich, MA, United States of America

Margaret Heider