THE USE OF CLASS IIB RESTRICTION ENDONUCLEASES IN 2ND GENERATION SEQUENCING APPLICATIONS

摘要

The present invention relates to a method for genotyping DNA molecules contained in at least one DNA sample comprising: (a) digesting the DNA molecules contained in at least one DNA sample with a class IIB restriction endonuclease to generate DNA fragments; (b) optionally separating DNA fragments comprising the recognition site for said class IIB restriction endonuclease from the remaining DNA fragments; (c) attaching at least one adaptor DNA to the 5' and/or 3' end of one or both strands of the DNA fragments comprising the recognition site for said class IIB restriction endonuclease obtained in a) or separated in b) to form adaptor-fragment constructs; (d) determining the sequence of at least a fraction of the DNA fragments obtained in c); and (e) assigning genotypes to said at least one DNA sample analysed based on the sequence data obtained in d). The present invention further relates to method for determining the position of DNA molecules comprised in a DNA library within the DNA sequence represented by said DNA library or within a known DNA sequence and for establishing a cross-reference between individual DNA molecules and their location in an at least three dimensional matrix.