The present disclosure provides, inter alia, specially designed DNA adaptors and methods of preparing the same. Methods and kits for carrying out and detecting marker-free precision genome editing and genetic variation using such adaptors are also provided.

The present disclosure provides, inter alia, specially designed DNA adaptors and methods of preparing the same. Methods and kits for carrying out and detecting marker-free precision genome editing and genetic variation using such adaptors are also provided.

The present invention provides a method for analysis of target nucleic acids which are present in low amounts. In particular, the method comprises the following steps: i. providing a sample wherein target nucleic acids are present in a low amount, ii. generating a reduced representation library of said target nucleic acids by a method comprising: fragmenting said target nucleic acids; ligating adaptors to said fragments; and selecting a subset of said adaptor-ligated fragments, iii. massively parallel sequencing said reduced representation library, and iv. identifying variants in said target nucleic acids by analyzing results obtained by said sequencing.

The present invention provides a method for analysis of target nucleic acids which are present in low amounts. In particular, the method comprises the following steps: i. providing a sample wherein target nucleic acids are present in a low amount, ii. generating a reduced representation library of said target nucleic acids by a method comprising: fragmenting said target nucleic acids; ligating adaptors to said fragments; and selecting a subset of said adaptor-ligated fragments, iii. massively parallel sequencing said reduced representation library, and iv. identifying variants in said target nucleic acids by analyzing results obtained by said sequencing.

The present disclosure provides improved detection (e.g., diagnostic) assays that utilize a Cas protein collateral cleavage activity.

The present disclosure provides improved detection (e.g., diagnostic) assays that utilize a Cas protein collateral cleavage activity.

Provided herein are methods, systems, and compositions for efficient nucleic acid assembly. Nucleic acid assembly may comprise assembly of variants comprising paired homology.

Provided herein are methods, systems, and compositions for efficient nucleic acid assembly. Nucleic acid assembly may comprise assembly of variants comprising paired homology.

It is a first object of the present invention to provide a method for preparing a Tile vector, being a vector, which comprises a selectable marker and a coding polynucleotide, wherein said coding polynucleotide is immediately preceded and followed by a type IIs recognition sequence, wherein said preceding and following recognition sequences are recognized by a same type IIs restriction enzyme, but have an opposite orientation. More particularly, the position and orientation of said preceding and following type IIs recognition sequences provides for the cleavage of said Tile vector by a corresponding type IIs restriction enzyme resulting in the release of said coding polynucleotide sequence having at its respective ends overhang sequences with a known orientation and length, while lacking said preceding and following type IIs recognition sequences. In a second object the present invention provides a method for using such Tile vectors obtained as previously described for joining two or more coding polynucleotides to form a product polynucleotide. Typically, said product polynucleotide is integrated in a vector.

It is a first object of the present invention to provide a method for preparing a Tile vector, being a vector, which comprises a selectable marker and a coding polynucleotide, wherein said coding polynucleotide is immediately preceded and followed by a type IIs recognition sequence, wherein said preceding and following recognition sequences are recognized by a same type IIs restriction enzyme, but have an opposite orientation. More particularly, the position and orientation of said preceding and following type IIs recognition sequences provides for the cleavage of said Tile vector by a corresponding type IIs restriction enzyme resulting in the release of said coding polynucleotide sequence having at its respective ends overhang sequences with a known orientation and length, while lacking said preceding and following type IIs recognition sequences. In a second object the present invention provides a method for using such Tile vectors obtained as previously described for joining two or more coding polynucleotides to form a product polynucleotide. Typically, said product polynucleotide is integrated in a vector.