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cDNA characterised using microarray technique, where cDNA are digested with restriction enzymes, then ligated and amplified by PCR and finally classified according to size to assist in human genome project
cDNA characterised using microarray technique, where cDNA are digested with restriction enzymes, then ligated and amplified by PCR and finally classified according to size to assist in human genome project
A method for characterising cDNA, so that DNA may be identified, for example, from a population of DNA, which method comprises: (a) Exposing a sample comprising a population of one or more cDNA or fragments thereof to a cleavage agent which recognises a predetermined sequence and cuts at a reference site at a known displacement from the predetermined sequence proximal to an end of each cDNA or fragment thereof so as to generate a population of terminal fragments; (b) Ligating to each reference site an adaptor oligonucleotide which comprises a recognition site for a sampling cleavage agent; (c) Exposing the population of terminal fragments to a sampling cleavage agent which binds to the recognition site and cuts at a sampling site of known displacement from the recognition site so as to generate in each terminal fragment a sticky end sequence of a predetermined length of up to 6 bases, and of unknown sequence; (d) Separating the population of terminal fragments into sub-populations according to sequence length; and (e) Determining each sticky end sequence. The method relates to a method for assaying DNA where the analysis of patterns of gene expression is required; especially analysis of gene expression in vivo.
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