New England Biolabs, Inc.

Other

Lixin Chen

Thomas C Evans, Jr

Chudi Guan

Barton E Slatko

Jennifer Ong

Romualdas Vaisvila

Ming-Qun Xu

Rebecca B Kucera

Laurence Ettwiller

Vladimir Potapov

Sriharsa Pradhan

Andrew F Gardner

Donald G Comb

Luo Sun

Henry P Paulus

Thomas C Evans

Rebecca Kucera

Romaldas Vaisvila

Inca Ghosh

Kelly M Zatopek

Andrew F Garner

Thomas Evans

Barton Slatko

Providing herein, among other things, is a method comprising incubating a double-stranded nucleic acid having a nick with a nick translating activity, a ligase, and a nucleotide mix comprising at least one modified nucleotide, to generate a product comprising a patch of a newly synthesized strand of a duplex nucleic acid containing a plurality of modified nucleoside monophosphates that are at or adjacent to the site of the nick. In some embodiments, the method may be used to map damaged nucleoside monophosphates in a nucleic acid. Compositions and kits for use in performing the method are also provided.

Mapping the location, type and strand of damaged and/or mismatched nucleotides in double-stranded DNA

Methods and compositions are provided for repairing a polynucleotide so that it can be copied with improved fidelity and/or yield in, for example, an amplification reaction. This involves the use of a reaction mixture that includes a ligase and a cofactor selected from NADor ATP and incubating the polynucleotide with the reaction mixture in the absence of Endonuclease VI. The reaction mixture may further contain an AP endonuclease and a polymerase. If used, these enzymes may be selected according to their ability to withstand high temperatures. For example, the reaction mixture may be used prior to a polynucleotide synthesis reaction in which case enzymes that are not thermophilic may be used. The repair reaction is not time sensitive with respect to seconds, minutes or hours of incubation in the enzyme mixture in as much as the repair is effected rapidly and prolonged incubation is not generally adverse.

Repair of nucleic acids for improved amplification

Methods and compositions are provided for repairing a polynucleotide so that it can be synthesized efficiently with improved fidelity and yield in, for example, an amplification reaction. This involves the use of a reaction mixture that includes a ligase and a cofactor selected from NAD+ or ATP and incubating the polynucleotide with the reaction mixture in the absence of Endonuclease VI. The reaction mixture may further contain an AP endonuclease and a polymerase. These enzymes are optionally selected according to their ability to withstand high temperatures so they can be included in an amplification mixture. The reaction mixture may be used prior to a polynucleotide synthesis reaction in which case enzymes that are not thermophilic may be used. The repair reaction is not time sensitive with respect to seconds, minutes or hours of incubation in the enzyme mixture.

A new type of transgene system is disclosed which allows efficient protein expression in a target host such as a plant, but avoids the undesirable result of the migration of the transgene into related host system and/or to the environment via the pollen. The methods described herein may also be applied to the expression of virtually any protein of interest (e.g. a toxic protein) in eukaryotic (yeast, insect, mammalian cells, etc.) and prokaryotic (, etc.) organisms.

Method for generating split, non-transferable genes that are able to express an active protein product

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Beverly, MA, United States of America

Lixin Chen